CD8A, GZMB, CCL5, CD2, and CXCL9, whereas CD8A, CD2, IL7R, CD27, and GZMB were hub genes of RA-SJ and OA group. The novel DEGs and functional pathways between RA and OA identified in this study may provide new insight into the underlying molecular mechanisms and therapeutic strategies of RA and OA.Rheumatoid arthritis (RA) and osteoarthritis (OA) are the most common joint disorders. Although they have shown analogous clinical manifestations, the pathogenesis of RA and OA are different. In this study, we used the online Gene Expression Omnibus (GEO) microarray expression profiling dataset GSE153015 to identify gene signatures between RA and OA joints. The relevant data on 8 subjects obtained from large joints of RA patients (RA-LJ), 8 subjects obtained from small joints of RA patients (RA-SJ), and 4 subjects with OA were investigated. Differentially expressed genes (DEGs) were screened. Functional enrichment analysis of DEGs including the Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were identified, which were mainly associated with T cell activation or chemokine activity. Besides, protein-protein interaction (PPI) network analysis was performed, and key modules were identified. Hub genes of RA-LJ and OA groups were screened, they were  According to recent studies, rheumatoid arthritis (RA) and osteoarthritis (OA) are considered to be the most prevalent rheumatic diseases, affecting 1% and 10% world’s population, respectively . RA is aPitx1, and functionally characterize 6 candidate human osteoarthritis genes  in mouse models  and was based on the platform of GPL570 Affymetrix Human Genome U133 Plus 2.0 Array [HGU133_Plus_2]. We collected samples from 24 subjects, including 10 small joints  from 10 RA patients, 10 large joints (knees) from the same 10 RA patients, and 4 OA subjects. The type of biological material used for gene expression quantification can be found in the study of Triaille C et al.  a |log2 (fold-change)| >1 and 2) an adjusted p < 0.05. Patients were divided according to their joint size. RA patients with large joints were compared with RA patients with small joints. R software was used to draw the volcano plot and PCA, HTML software was used to create the heatmap for the DEGs, and Venn diagrams of DEGs were drawn using Venny 2.1 (https://bioinfogp.cnb.csic.es/tools/venny/index.html), and uniform manifold approximation and projection (UMAP) were performed via R’s “umap” package , functional enrichment analyses of DEGs were conducted, including Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. A GO analysis consists of three biological processes (BP), a cellular component (CC), and a molecular function (MF), which provide a framework for describing the functions of gene products in all organisms. We assigned DEGs to specific pathways using KEGG pathway analysis (15). The threshold for significance was set at Benjamini-adjusted p < 0.05 and an enriched gene count of at least five genes enriched in the pathway.Using Enrichr (https://string-db.org/), a web-based database dedicated to predicting protein-protein interactions consisting of both physical and functional relations. We used Cytoscape v3.8.2 software to visualize and build the PPI network simultaneously while mapping the DEGs onto the PPI network with a medium confidence score of 0.4. Using ClusterOne from Cytoscape’s software suite, the gene network clustering analysis was conducted to refine the key PPI network modules. An R software package with the “ggplot2” package was used to analyze the expression levels of key PPI network modules , anti-CD2 antibodies , anti-GZMB antibodies , anti-CCL5 antibodies , anti-CXCL9 antibodies , anti-IL7R antibodies , and anti-CD27 antibodies . Incubation at room temperature for 30 min was followed by diluted biotinylated secondary antibodies. After incubation in Street Avidin-Biotin Complex for 20 min, sections were treated with diaminobenzidine (DAB) substrate solution until the desired color intensity was achieved . Images p-value, the Benjamini–Hochberg FDR  was used. Hypergeometric tests were used to distinguish significantly enriched GO terms and KEGG pathways. Statistical analysis of significant differences was conducted using one-way ANOVA by Prism 7 software. A p-value less than 0.05 was considered statistically significant.To correct the versus OA and RA-SJ versus OA, respectively; versus the OA group and 62% of downregulated genes in the RA-LJ group versus the OA group.The characteristics of the samples were listed in versus OA groups are mainly T cell receptor complexes in the CC category, chemokine receptor binding in the MF category, and T cell activation in the BP category. The T cell receptor signaling pathway is the most enriched KEGG pathway. The DEGs in RA-SJ compared to OA groups include collagen-containing extracellular matrix components, protein homodimerization activity in MF, and positive regulation of B cell activation and cytokine-mediated signaling pathways in BP. Pathways associated with hematopoietic cells were identified as the most significantly altered pathways in RA-SJ compared to OA.To take a further step in the investigation of the biological functions of DEGs, a functional enrichment analysis was conducted and the results are presented in CST7, CD8A, GEMK, GZMH, GZMA, GZMB, PRF1, CD3D, CXCL9, NKG7, CD2, CD247, ITGAL, CCL5) and three downregulated genes . While in versus OA.In accordance with the STRING web-based database, two PPI networks were built with an interaction score >0.4 . In ordeCD8A, GZMB, CCL5, CD2, and CXCL9, whereas CD8A, CD2, IL7R, CD27, and GZMB. Among them, CD8A, GZMB, and CD2 are hub genes in both groups. In CD8A, CD2, GZMB, CCL5, CXCL9, IL7R, CD27) were shown. In RA joints, CD8A, CD2, GZMB, CCL5, CXCL9, IL7R, and CD27 levels were significantly higher than in OA joints. Further, IHC was used to determine the expression levels of seven different hub genes in large and small joints of RA and OA joints. As shown in Based on their degree value, which represents the number of interactions among the genes, the hub genes were sorted. versus OA groups, CD8A, GZMB, CCL5, CD2, and CXCL9 are the hub genes, while CD8A, CD2, IL7R, CD27, and GZMB are the hub genes in RA-SJ versus OA groups. Lastly, immunohistochemistry was used to validate the protein expression levels of the 7 hub genes in RA and OA joint tissues.Rheumatoid arthritis and osteoarthritis are two of the most common of the more than 100 different types of arthritis. As a result of chronic inflammation and autoimmunity associated with RA disease, synovial fluid is produced by the membrane lining, causing synovitis, joint pain, and eventually chronic and progressive erosion . As a whBased on our analysis, we found that GO enrichments were mainly related to T cell activation, B cell activation, chemokine activity, and collagen-containing extracellular matrix in large and small joints of RA. In RA joints, T cell receptor signaling pathways and cytokine-cytokine receptor interactions were the main signaling pathways. It is easy to understand from these results for that rheumatoid arthritis is an autoimmune and inflammatory disease, which also suggests different treatment strategies for RA and OA. Rituximab, a kind of anti-CD20 chimeric monoclonal antibody that can inhibit the proliferation of stimulated human B cells, has long been a choice for refractory rheumatoid arthritis . MoreoveCCL5, also known as RANTES, its protein is a chemoattractant for blood monocytes, memory T helper cells, and eosinophils. It causes the release of histamine from basophils and activates eosinophils and plays an active role in the chemotactic activity of T cells in RA by facilitating leukocyte infiltration (CXCL9, whose protein is detected in sera, synovial fluid, and synovial tissue (Hub genes ltration . Solomonltration . The patl tissue . By bind+ T cells from patients with RA transition into highly proliferative, tissue-invasive and proinflammatory effector cells (+ T cells rapidly induce synovitis in a human synovium mouse chimera model (+ T cells characterized by GZMK+, GZMB+, and GNLY + phenotypes in RA joint synovial tissues by integrating single-cell transcriptomics and mass cytometry (GZMB in the classification of CD8+ T cells in RA. It has already been found for decades that there was a remarkable increase in GAMB concentrations in plasma and synovial fluid of patients with established rheumatoid arthritis compared with disease controls (+ T cell numbers are elevated and correlate with the levels of rheumatoid factor, supporting a role for CD27 in human RA (−CD70 interactions with anti-CD70 antibody reduces bone and cartilage erosion and inflammatory infiltrates in the joints of mice with collagen-induced arthritis, moreover, decreases collagen-specific antibody production (T cells participate in multiple pathways driving the disease process in rheumatoid arthritis. Naive CD4or cells . Equippera model . It was ra model . To dateytometry , indicatcontrols . Besidescontrols . In addicontrols . It has human RA . Blockinoduction . In summCD8A, CD2, GZMB, CCL5, CXCL9, IL7R, and CD27. As a result of our study, we may be able to identify new diagnostic markers and therapeutic targets for RA and OA.In conclusion, we screened differentially expressed genes between large and small joints of RA between OA joints in this study. Several functional and pathway enrichments were also found between RA and OA joints, primarily related to T cell activation or chemokine activity. It was determined whether RA and OA joint samples expressed hub genes such as


DGAT1, DGAT2, or SEIPIN, which were involved in TG synthesis. However, it did reduce the expressions of FITM2, CIDEA, and FSP27, which were major involved in lipid droplet growth and budding, and lipid droplet expansion. In addition, exercise reduced ATGL protein level in HFD mice, and regulated lipophagy-related markers, including increasing ATG5, LAMP1, LAMP2, LAL, and CTSD, decreasing LC3II/I and p62, and promoting colocalization of LAMP1 with LDs. In summary, our data suggested that 15 weeks of moderate treadmill exercise was beneficial for regulating liver lipid droplet dynamics in HFD mice by inhibiting abnormal lipid droplets expansion and enhancing clearance of lipid droplets by lysosomes during the lipophagic process, which might provide highly flexible turnover for lipid mobilization and metabolism. Abbreviations: β-actin: actin beta; ATG5: autophagy related 5; LAMP2: lysosomal-associated membrane protein 2; LAMP1: lysosomal-associated membrane protein 1; SQSTM1/p62: sequestosome 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; ATGL: adipose triglyceride lipase; CSTD: cathepsin D; LAL: lysosomal acid lipase; DGAT1: diacylglycerol-o-acyltransferase 1; DGAT2: diacylglycerol-o-acyltransferase 2; CIDEA: cell death inducing dffa-like effector a; CIDEC/FSP27: cell death inducing dffa-like effector c; FITM2: fat storage-inducing transmembrane protein 2; PLIN2: adipose differentiation related protein; PLN3: tail-interacting protein 47; HSP90: heat shock protein 90; SREBP1c: sterol regulatory element binding protein-1c; chREBP: carbohydrate response element binding protein. Lipid droplet is a dynamic organelle that undergoes periods of biogenesis and degradation under environmental stimuli. The excessive accumulation of lipid droplets is the major characteristic of non-alcoholic fatty liver disease (NAFLD). Moderate aerobic exercise is a powerful intervention protecting against the progress of NAFLD. However, its impact on lipid droplet dynamics remains ambiguous. Mice were fed with 15 weeks of high-fat diet in order to induce NAFLD. Meanwhile, the mice performed 15 weeks of treadmill exercise. Our results showed that 15 weeks of regular moderate treadmill exercise alleviated obesity, insulin intolerance, hyperlipidemia, and hyperglycemia induced by HFD. Importantly, exercise improved histological phenotypes of NAFLD, including hepatic steatosis, inflammation, and locular ballooning, as well as prevented liver fat deposition and liver injury induced by HFD. Exercise reduced hepatic lipid droplet size, and moreover, it reduced PLIN2 protein level and increased PLIN3 protein level in the liver of HFD mice. Interestingly, our results showed that exercise did not significantly affect the gene expressions of  Non-alcoholic fatty liver disease (NAFLD) has emerged as the primary cause of chronic liver diseases worldwide, with an estimated global prevalence of 25% . NAFLD pImportantly, an unhealthy lifestyle, which includes factors such as saturated fat-rich diets, cholesterol fat-rich diets, sugared drinks as well as lack of physical activity, has been directly related to the susceptibility of developing NAFLD . CurrentEven though regular bouts of aerobic exercise training are considered beneficial for decreasing hepatic lipid content , it may Therefore, the aim of this study was to investigate the effect of 15 weeks of regular treadmill exercise on preventing NAFLD. Moreover, we aim to characterize the potential mechanism of LD dynamics, including LD biogenesis and degradation. We found that exercise could regulate the size and content of hepatic LDs by inhibiting the abnormal expansion of LDs and restoring lipophagic activity, which might provide a flexible dynamic process for lipid mobilization and metabolism.In this case, 24 C57BL/6 male mice (5 weeks of age) were purchased from the animal experiment center of East China Normal University . All the mice were housed in a specific pathogen-free room with conventional laboratory conditions  on a 12:12 h light-dark cycle (6 a.m.–6 p.m.), with access to water and food ad libitum. Body weight was measured weekly until the mice were sacrificed.n = 6), Normal diet combined exercise , High-fat diets sedentary , and High-fat diet combined exercise . The mice from HFD groups (HS and HE) were fed with a high-fat diet contained 60% fat, 20% carbohydrate, and 20% protein in terms of kcal  for 15 weeks , 60 min for formal training (13 m/min), and 5 min for relaxing (7 m/min). Mice from Group NS and Group HS were also placed on the static treadmill for the same duration of time. We avoided using electric shock stimulation in our experiments and used a cork stick to push and drive the mice throughout to reduce their pain. Their body composition  was analyzed 24 h after the last exercise intervention. The mice were sacrificed 36 h after the last training session, following overnight fasting.One week before the end of the exercise intervention, ITT was carried out. After 4 h of fasting (9:00 a.m. to 1:00 p.m.), blood was collected from the tails for glucose measurement at the time of 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min. After the first blood glucose test (t = 0 min), the mice immediately accepted an injection  of insulin  at 0.5 U/kg body weight.The blood samples were centrifuged  at 3000 r/min for 10 min to acquire the serum. Serum samples were stored at −80 °C. All the assay kits were obtained from Nanjing Jiancheng Bioengineering Institute  for serum TC (A111-1-1), LDL-c (A113-1-1), aspartate aminotransferase (AST) (C010-2-1), and alanine aminotransferase (ALT) (C010-2-1).n = 3). Evaluation of NAFLD activity score (NAS) has been described before [The liver tissues were extracted immediately after harvesting and immersed in 10% neutral buffered formalin for 24 h. One part of these tissues was dehydrated using 70% ethanol, embedded in the paraffin, and cut into a 5 μm coronal section for hematoxylin and eosin. The same liver lobe of each animal was captured by using the Nikon View software   and cut into a 5 μm coronal section for Oil Red O staining. The images were visualized under light microscope . The number and area of LDs were analyzed with Image J version 1.51 software and Image-pro plus 6.0 . All of these features were scored in a blinded manner per sample.The total RNA was extracted from liver samples (40 mg per sample) by the Trizol method , as per the manufacturer’s instruction. For real-time PCR analysis, reversing transcription of RNA was performed using a cDNA reverse transcription kit . Gene expression levels were further detected by using an RT-PCR reaction mixture containing SYBR-green fluorescent dye . Primers used are listed in g. The supernatant was collected. Bradford method (P0010S) was used to determine the protein concentration of total homogenate.Western blot was conducted as previously described . Liver tThe samples (25 µg) were separated on 10–12% SDS-PAGE gel and transferred to the PVDF membrane (Millipore). The membranes were then blocked in QuickBlock™ Blocking Buffer for Western Blot  for 1.5 h at room temperature and incubated with the following primary antibodies: ADRP/perilipin2 (15294-1-AP), LC3B (14600-1-AP), lysosomal acid lipase (LAL)(12956-1-AP), and Cathepsin D (CTSD)(21327-1-AP) were obtained from Proteintech , adipose triglyceride lipase (ATGL)(#2439), Beclin1 (#3495S), ATG5 (#12994), and LAMP1 (#3243) were obtained from Cell signaling technology , TIP47/PLIN3 , anti-sqstm1/p62 , LAMP2 , and horseradish peroxidase-conjugated secondary antibodies . The images of protein bands were captured and analyzed by the ChemiDoc MP Imaging System . The β-actin , HSP90 , alpha-tubulin , and GAPDH  were used to normalize the amount of protein loaded. Unless otherwise specified, all biochemical reagents were purchased from Beyotime, Shanghai, China.The paraffin-embedded liver tissue sections were cut into 10 μm coronal sections and used for immunofluorescence staining. These prepared sections were blocked with mouse serum and incubated with ADRP/perilipin2  overnight at 4 °C, followed by fluorescence conjugated secondary antibody . The sections then added CY3-TSA  and were incubated in dark for 10min, followed by antigen retrieval with EDTA antigen retrieval buffer . Subsequently, these sections were incubated with the second primary antibody LAMP1 , and followed by a fluorescence conjugated secondary antibody from Servicebio. The immunofluorescence was captured by using a Nikon Eclipse C1 microscope.p  <  0.05. All descriptive values are reported as means ± SEM.Statistical analysis was performed using GraphPad Prism . A two-way analysis of variance (ANOVA) with Turkey methods (with high-fat diet and exercise intervention as factors) was used for the evaluation of differences among the four groups. ITT data were analyzed using repeated-measures ANOVA. The Pearson correlation coefficient was used to calculate correlation coefficient. The statistical significance was considered at Poor dietary habits are highly associated with the development of NAFLD . In our NAFLD, recognized as a hepatic manifestation of metabolic syndrome, is usually accompanied by insulin resistance, hyperglycemia, and dyslipidemia as background factors . We firsAbnormal hepatic LDs deposition is the main characteristic of NAFLD, which may further lead to liver injury . In thisHistological examination of liver tissues revealed that exercise significantly reduced the score of hepatic steatosis, inflammation, locular ballooning, and NAFLD activity in HFD mice D,E. The As the above results showed, exercise is sufficient to decrease the accumulation of hepatic LDs in HFD mice. With this in mind, we next investigated the molecular mechanism of exercise in regulating hepatic LDs dynamics. In general, LDs accumulation is coordinated by transformation of LD membrane proteins and accumulation of lipid core .As shown in In order to investigate the mechanism that exercise regulated hepatic LDs biogenesis, the expression levels of genes involved in controlling triglycerides synthesis, LD growth and budding, and LD expansion, including DGAT1, DGAT2, FITM2, SEIPIN, CIDEA, and FSP27, were examined D. The reLipolysis and lipophagy are two major forms of LD degradation within cells. In order to evaluate the degradation pathway of LDs, we first measured ATGL, the key enzyme in the first step of neutral lipid lipolysis . Our resNotably, lipophagy is a special form of autophagy. During this process, LDs were sequestered in autophagosomes (lipophagosomes) for degradation in lysosomes. In general, the LD sequestered in the lysosome is a symbol of lipophagy . Here, tGiven that lysosomes are considered as the destination for lipophagy, we further evaluated protein levels of molecules involved in sustaining hepatic lysosomal function, including LAMP1, LAMP2, CTSD, and LAL. The results showed that HFD induced a decrease in protein levels of LAMP2 and LAL, and an inhibition in CTSD maturation (increased pro-CTSD/mature-CTSD). On the contrary, exercise significantly increased the protein levels of LAMP1, LAMP2, and LAL, and decreased the protein levels of pro-CTSD/mature-CTSD in HFD mice H–L.Our findings demonstrated that LD dynamics was an important hepatic mechanism of exercise benefits during the development of NAFLD. Our data showed that 15 weeks of HFD tilted the balance of LD dynamics towards increased biogenesis , and thus resulted in large-size LD accumulation. At the same time, the degradation of LDs was partly inhibited. On the contrary, 15 weeks of moderate treadmill exercise inhibited the unorganized LD expansion and titled the balance of LDs dynamics towards lipid degradation via lipophagy.Clinical evidence shows that subjects with NAFLD are more likely to have a greater body mass index, higher serum concentrations of total cholesterol, increased fasting glucose level, and insulin resistance . In thisAs expected, we observed that 15 weeks of exercise intervention in HFD mice was sufficient for preventing the development of NAFLD, manifested by alleviation of HFD-induced obesity, insulin intolerance, abnormally elevated fasting glucose, serum TC and LDL-c, liver weight, liver triglyceride, NAFLD activity score, and increased liver damage.It is well known that LD is the majority form of triglyceride storage. Therefore, we further evaluated hepatic LD content. Our results showed that HFD induced an abnormal increase in content, area, and number of hepatic LD, which was ameliorated by 15 weeks of aerobic exercise intervention D–H. The LDs undergo dynamic periods of biogenesis and degradation under different environmental stimuli, such as nutrient deprivation, and hypoxia. The PAT family proteins, including perilipin (PLIN1), ADRP (PLIN2), TIP47 (PLIN3), S3-12 (PLIN4) and OXPAT (PLIN5), have been shown to play an important role in regulating LD dynamics. In mammals, PLIN2, located almost exclusively in LDs, is closely associated with intracellular lipid content, and is highly expressed in the liver of fatty liver patients . PLIN3 A,B. MoreTo further confirm and understand this phenomenon, we measured the expression of genes involved in several steps of LD biogenesis, such as TG synthesis, LD growth and budding, and expansion of LDs E–J. ConsThe change In the pattern of lipid droplet accumulation induced by exercise might provide a higher chance for energy mobilization, since inhibition of unlimited fusion and expansion of LD increases the total surface of the LDs and thereby increases the access of several lipid degradation components to stored lipid . In geneLipophagy exerts physiological functions by eliminating LD via the autophagic core mechanism . In our −/− mice by gene or enzyme methods could decrease the abnormal excess lipid storage in the liver [We, thus, further measured lipophagy by performing immunofluorescence staining. In this case, 15 weeks of exercise intervention was able to restore the HFD-induced impaired colocalization of PLIN2 and LAMP1 G. Consishe liver ,50. In t


ETV4 fusion in prostate cancer are not fully understood, and drugs targeting ETV4 fusion have not been developed. To examine key cellular signaling pathways and explore therapeutic targets and drugs for ETV4-fusion-positive prostate cancer, we analyzed RNA sequencing data and clinical information for prostate cancer. The ETV4-fusion-positive group was selected through prior study and analysis comparing ETV4-fusion-positive and -negative groups was conducted using a Pearson correlation test. We obtained 393 genes correlated with ETV4 expression. Pathway analysis was performed using over-representation analysis (ORA), and six cancer-specific molecular signaling pathways  were altered in the ETV4-fusion-positive group. Furthermore, a gene–drug database was used to find an actionable drug and therapeutic target for the ETV4-fusion-positive group. Here, we have identified significantly altered genes and oncogenic signaling pathways in ETV4-fusion-positive prostate cancer, and we suggest therapeutic targets and potential drugs for ETV4-fusion-positive prostate patients.Gene fusion between androgen receptor (AR) response genes and E26 transformation-specific (ETS) family members increases the gene expression of ETS family members, and promotes tumorigenesis in prostate cancer. However, the molecular features of  Prostate cancer is the second-most frequent cancer type in males following lung cancer, and it ranked as the fifth-leading cause of death in males in 2020 . It is aTMPRSS2. Prostate cancer frequently harbors the chromosomal rearrangement of the E26 transformation-specific (ETS) gene, and the most frequent fusion partner gene is ERG and TMPRSS2 fusion is found in about 50% of prostate cancers [ETV1 and TMPRSS2 fusion is found in 5% of prostate cancers. Fusion of the ETS gene results in the high expression of ERG or ETV1, and it is an important cause of prostate tumorigenesis and tumor development [ETV4, ETV5, and SKIL have been discovered in studies [ETV4), defined as another driver mutation, was observed in 2% of prostate cancers [TMPRSS2 is a major fusion companion of ETV4, and a significant portion fuses by gene-intergenic fusion, an unusual mechanism which produces chimeric RNA [STAT3 and PMEPA1, are also known to fuse with ETV4. Also, there are some cases where the mechanism by which ETV4 is highly expressed in prostate cancer is not well-defined. The overexpression of ETV4 promotes cell proliferation by upregulating cancer-related genes and the epithelial–mesenchymal transition (EMT) by activating EMT-specific transcription factors [ETV4 increases metastasis by activating PI3K kinase-RAS signaling in mouse prostate cancer models [ETV4 are well-described in both in vivo and in vitro models [ETV4-fusion-positive prostate cancers have not been fully elucidated. Indeed, therapeutic targets and candidate targeted drugs for ETV4-overexpressing prostate cancer remain unknown. Thus, we here systematically analyzed RNA sequencing (RNA-seq) data obtained from The Cancer Genome Atlas (TCGA) database and identified oncogenic signaling pathways activated in ETV4-fusion-positive prostate cancer, which results in ETV4 overexpression. We further performed gene–drug network analysis and investigated therapeutic targets and candidate drugs in ETV4-fusion-positive prostate cancer.elopment ,4. In ad cancers . TMPRSS2eric RNA . Other g factors . ETV4 inr models . Overallo models . HoweverETV4-fusion-positive samples were selected as the case group, and 86 samples without any driver mutations, such as ERG fusion, ETV1 fusion, SPOP mutation, FOXA1 mutation, or IDH1 mutation, were selected as the control group.Structural variation, mutation information, and gene expressions from The Cancer Genome Atlas (TCGA) prostate cancer molecular taxonomy study was used to clarify subtypes of prostate cancer . A totalhttp://gdac.broadinstitute.org, accessed on 12 December 2021). Sample type code 01, indicating primary solid tumor samples, and sample type code 11, indicating adjacent normal tissue, were used in this study. ETV4 fusion with androgen-responsive genes results in ETV4 overexpression in prostate cancers [ETV4 expression via the Pearson correlation test using an absolute value of Pearson correlation coefficient over 0.3 (|R| > 0.3), assuming that the expression fluctuation of downstream genes affected by ETV4 would be proportional to the expression of ETV4. RNA expression data with upper quantile normalization was obtained from the TCGA Prostate Adenocarcinoma (PRAD) study from the Broad GDAC Firehose website  was used to identify cancer-related genes [The CancerMine database  via ConsensusPathDB  with the options of allowing a minimum of two genes to overlap from the input list and a p-value < 0.01 cutoff. Biocarta, Ehmn, Humancyc, Inoh, Kegg, Manual upload, Netpath, Pharmgkb, Pid, Reactome, Signalink, Smpdb, and Wikipathways, which were curated and provided by CPDB, were used for pathway analysis. The genes significantly correlated with https://civicdb.org/home, accessed on 22 June 2021). Gene–drug network analysis was performed using genes involved in cancer-specific pathways of ETV4-fusion-positive prostate cancer, and therapeutic targets and potential actionable drugs were obtained via the network analysis.A drug-target database containing target genes, variants, disease, and drugs was obtained from the Clinical Interpretation of Variants in Cancer platform  in software R version 3.6.3. Because the clinical information provided from TCGA is limited, only the available data sets were used for this statistical analysis. Clinical characteristics, such as age, prostate-specific antigen (PSA) level, Gleason score, tumor stage (T stage and M stage), laterality, and ethnicity were analyzed between the ETV4 expression were identified via a Pearson correlation test using the cor.test function in R. Cancer-specific cellular signaling pathways and involved genes altered in ETV4-fusion-positive prostate cancer were visualized at the gene level using an R package named ComplexHeatmap [Genes significantly correlated with xHeatmap . Graphs,xHeatmap .ETV4-fusion-positive prostate cancers were selected based on structural variation data and gene expression data according to a previous report [ETV4-fusion-positive prostate cancers and 86 ETV4-fusion-negative prostate cancers were analyzed. Age, PSA level, Gleason score, tumor stage (T stage and M stage), laterality, and ethnicity were compared between the ETV4-fusion-positive and -negative prostate cancer groups, and no significant difference was observed  was applied to identify genes significantly correlated with ETV4 expression. A total of 393 genes were obtained, and interestingly, except for 10 genes, most were positively correlated  and a negative correlation between ETV1/ETV4 and PXN (R = −0.3) were observed. Moreover, PXN expression in ETV4-fusion-positive prostate cancer was lower compared to normal (A total of 85 genes were significantly highly expressed, and only e cancer . Paxillie cancer ,16,17. P cancers . Consisto normal . CYP1B1, CYP3A4, HSPA5, RAN, SULT2A1, and TOP1, were involved in more than two pathways, and multiple targeted pathways included the tamoxifen pathway, metabolism, the irinotecan pathway, and androgen receptor signaling. Cellular signaling pathways altered in ETV4-fusion-positive cancer were validated by analyzing microarray data from a previous study [ETV4 in a prostate cancer cell line, and signaling pathways were analyzed using these genes. When ORA was performed using 774 genes, metabolism, interferon signaling, and MAPK/NF-kB signaling, among seven inferred pathways, were consistent with the results of this study using the TCGA database (Gene-pathway network analysis was further performed . Six genus study . In the database .ETV4-fusion-positive prostate cancer, gene–drug network analysis was performed using 86 genes involved in cancer-specific pathways of ETV4-fusion-positive prostate cancer. PARP1, NQO1, HSPA5, and TOP1 were identified as potential molecular targets for ETV4-fusion-positive prostate cancer, and olaparib, amrubicin, fluorouracil, and irinotecan were suggested as candidate drugs, respectively (HSPA5 and TOP1 were associated with the irinotecan pathway and androgen receptor signaling, while PARP1 was categorized under the androgen receptor signaling pathway, and NQO1 was related to metabolism (To suggest therapeutic molecular targets and candidate drugs for ectively . HSPA5 atabolism .ETV4 rearrangement has lately been discovered as a driver gene in 2% of prostate cancers. The small number of ETV4 subtypes among prostate patients and the short history of research on ETV4 rearrangement have limited understanding of the molecular features of ETV4 subtypes of prostate cancer and, in turn, have resulted in an absence of effective targeted therapy for these patients. In addition, since ETV4 is a transcription factor, it is difficult for it to be a direct drug target, unlike proteins that have a kinase domain or cell-surface receptor [ETV4 rearrangement is mutually exclusive with other oncogenic driver mutations in prostate cancer, the study of the molecular features, specific therapeutic targets, and potential drugs for ETV4 subtypes of prostate cancer are needed. In this study, we first identified 393 genes correlated with ETV4 expression and six cancer-specific cellular signaling pathways, namely the irinotecan pathway, metabolism, androgen receptor signaling, interferon signaling, MAPK/NF-kB signaling and the tamoxifen pathway, altered by the presence or absence of ETV4 fusion. These signaling pathways could be further categorized into three groups: hormone-related pathways, metabolic pathways, and inflammation/cancer pathways. ERG subtypes of prostate cancer have altered androgen receptor signaling, similar to the ETV4 subtypes of prostate cancer shown in the current study [ERG subtypes of prostate cancer [ETV4-fusion-positive prostate cancer, based on its unique features of cellular signaling pathways. Estrogen treatment was once suggested as an alternative method to suppress testosterone in prostate cancer [ETV4-fusion-positive prostate cancer, and all genes related to the tamoxifen pathway were positively correlated with ETV4. These observations suggest the advantage of using hormonal therapy in ETV4-fusion-positive prostate cancer. As half of the genes included in the six cancer-specific pathways were related to the metabolism pathway, interestingly, the metabolic pathway was highly associated with the ETV4 subtype of prostate cancer (ETV4-fusion-positive cancer. MAPK/NF-kB signaling was also altered in the ETV4 subtype of prostate cancer (Androgen signaling is stimulated by androgen, and activated androgen signaling multiplies and spreads prostate cancer cells . Anti-annt study . A previe cancer , and we e cancer , and cone cancer . Metabole cancer . The MAPETV4-fusion-positive prostate cancer. Genes associated with ETV4-fusion-positive prostate cancers were used as the input, and four therapeutic candidate targets, PARP1, NQO1, HSPA5, and TOP1, and the respective selective drugs, olaparib, amrubicin, fluorouracil, and irinotecan, were identified. Olaparib, the first targeted drug for prostate cancer, has recently been approved for BRCA-mutated metastatic castration-resistant prostate cancer [ETV4 subtype of prostate cancer, regardless of BRCA mutation. A large clinical study should be warranted for evaluation of the effects of olaparib on the ETV4 subtype of prostate cancer in the future. Next, we performed gene–drug network analysis and identified new therapeutic targets and drugs for e cancer . In addiPXN, which is known as an oncogene in prostate cancer [ERG and SPOP subtypes of prostate cancer, as compared to normal cells, and lower expression in ETV1 and ETV4 subtypes of prostate cancer, as compared to normal cells (PXN is known to be related with tumor progression, invasion, and metastasis via EMT [ERG fusion or SPOP mutation could share this cancer progression mechanism while a prostate cancer subtype with an ETV4 fusion or ETV1 fusion does not.Furthermore, we found different gene expression patterns in different types of prostate cancer. For example, e cancer , has higal cells . This sh via EMT ,32,33,34ETV4-fusion-positive prostate cancer patients were systematically identified. Cancer genomic data, cancer transcriptomic data, clinical data, and patient information were obtained from the TCGA database. Microarray data performed in cell lines after upregulating or downregulating ETV4 was collected from prior study data and utilized to conclude the molecular function of ETV4. The future validation of the potential therapeutic targets and potential drugs in in vitro or in vivo models would demonstrate the efficiency of our approach.In this study, cellular signaling pathway analysis was extensively performed, and novel therapeutic targets and candidate drugs for ETV4 overexpression was observed in various cancer types, such as prostate cancer, bladder cancer, gastric cancer, colon cancer, and hepatocellular carcinoma, and the underlying mechanism and oncogenic function have been revealed, which suggest the importance of ETV4 in cancer research [ETV4-associated human cancers.research . TherefoETV4 subtype of prostate cancer. To suggest potential therapeutic targets and actionable candidate drugs, gene–drug network analysis was performed, and four genes, HSPA5, NQO1, PARP1, and TOP1, and their targeted drugs, fluorouracil, amrubicin, olaparib, and irinotecan, respectively, were identified. In this study, we discovered that six cancer-specific cellular signaling pathways, the irinotecan pathway, metabolism, androgen receptor signaling, interferon signaling, MAPK/NF-kB signaling, and the tamoxifen pathway, were altered in the ETV4-fusion-positive prostate cancer. Considering the absence of targeted drugs for the ETV4 subtype of prostate cancer, our study will allow a step forward in investigations of treatments. Furthermore, the analysis algorithms we established in this study could be applied to analyze the molecular features of other types of cancer, and in turn, this could provide enormous benefits in developing new drug treatments in regard to cost, time, and the prediction of adverse effects.Taken together, we have provided information on altered cellular signaling pathways and therapeutic targets for


The ketone bodies (KBs) β-hydroxybutyrate and acetoacetate are important alternative energy sources for glucose during nutrient deprivation. KBs synthesized by hepatic ketogenesis are catabolized to acetyl-CoA through ketolysis in extrahepatic tissues, followed by the tricarboxylic acid cycle and electron transport chain for ATP production. Ketogenesis and ketolysis are regulated by the key rate-limiting enzymes, 3-hydroxy-3-methylglutaryl-CoA synthase 2 and succinyl-CoA:3-oxoacid-CoA transferase, respectively. KBs participate in various cellular processes as signaling molecules. KBs bind to G protein-coupled receptors. The most abundant KB, β-hydroxybutyrate, regulates gene expression and other cellular functions by inducing post-translational modifications. KBs protect tissues by regulating inflammation and oxidative stress. Recently, interest in KBs has been increasing due to their potential for treatment of various diseases such as neurological and cardiovascular diseases and cancer. Cancer cells reprogram their metabolism to maintain rapid cell growth and proliferation. Dysregulation of KB metabolism also plays a role in tumorigenesis in various types of cancer. Targeting metabolic changes through dietary interventions, including fasting and ketogenic diets, has shown beneficial effects in cancer therapy. Here, we review current knowledge of the molecular mechanisms involved in the regulation of KB metabolism and cellular signaling functions, and the therapeutic potential of KBs and ketogenic diets in cancer. Ketone bodies (KBs) are a group of water-soluble molecules that contain the ketone groups, which consist of acetoacetate (AcAc), D-β-hydroxybutyrate (BHB), and acetone. Among KBs, BHB is the most abundant KB, accounting for about 70% of the circulating KB pool ,2. BHB aKBs are produced by ketogenesis mostly in hepatocytes, but also to a minor extent in astrocytes, cardiomyocytes, renal epithelia, and enterocytes ,6,7. Aftketonemia, and a condition in which excretion in the urine is increased is called ketonuria. In general, ketonemia and ketonuria are collectively referred to as ketosis. However, in certain pathological conditions, such as diabetic ketoacidosis, KBs can exceed 20 mM. Under physiological conditions, plasma concentrations of KBs in adult humans range from 0.05–0.1 mM, rising to 1–2 mM after 2 days of fasting, and levels reaching 5–8 mM under prolonged fasting and starvation ,12. The Although diabetic ketoacidosis is a pathological condition, mild ketonemia  resulting from fasting, a KD, or other dietary interventions including calorie restriction (CR) has been demonstrated to be beneficial, improving metabolic profiles and extending healthy lifespan ,14,15,16Cancer cells undergo metabolic reprograming to maintain rapid cell proliferation and tumor growth ,28,29. AThe pathways of hepatic ketogenesis and extrahepatic ketolysis are shown in The ketone bodies AcAc and BHB are released into the circulation and taken up by extrahepatic tissues through MCTs. Ketolysis occurs in the extrahepatic mitochondria, where BHB is converted to AcAc by BDH1 and acetyl-CoA through a reaction catalyzed by succinyl-CoA:3-oxoacid-CoA transferase (SCOT) and ACAT1 . After aHMGCS2 gene expression and activation of HMGCS2 protein, a key rate limiting enzyme in ketogenesis [Ketogenesis is regulated at several levels and the main regulatory points for ketogenesis include mobilization of free fatty acid, fatty acyl-CoA transport into mitochondria, and ogenesis ,6. Mobilogenesis ,45. FattHMGCS2 gene expression is regulated at the transcriptional level and the activity of HMGCS2 protein is regulated at the post-translational level [HMGCS2 gene transcription is regulated by insulin and glucagon ratios mediated by the transcription factor forkhead box A2 (FOXA2) [HMGCS2 transcription by FOXA2 acetylation via the cAMP-p300-FOXA2 pathway, which is inhibited upon deacetylation of FOXA2 by the NAD+-dependent enzyme sirtuin 1 (SIRT1) [al level ,6,46. Re (FOXA2) . Insulin (FOXA2) . In cont (SIRT1) . HMGCS2 gene expression is positively regulated by the peroxisome proliferator-activated receptor α (PPARα) [HMGCS2 gene expression in the liver through the expression of fibroblast growth factor 21 (FGF21), known as an endocrine regulator of ketotic states [ (PPARα) . Recent c states ,6,50,51.HMGCS2 transcription is negatively regulated by mechanistic target of rapamycin complex 1 (mTORC1) kinase [Meanwhile, ) kinase ,53 and t) kinase , presuma) kinase ,56.HMGCS2 gene expression is positively regulated by PPARγ, which is inhibited by the Wnt/β-catenin signaling pathway in intestinal epithelial cancer cells [Recently, it has been reported that er cells . SLC38A4er cells . The HMGCS2 protein undergoes several different types of post-translational modifications, as shown in Oxct1 and Oxct2 [Oxct1 mRNA levels in most cell lines were higher than those of the testis-specific isoform, Oxct2, suggesting that Oxct1 may play a more dominant role than Oxct2 in ketolysis [Oxct1 gene product, OXCT1, is abundant in the brain, heart, and kidney, and can be detected in almost all extrahepatic tissues [Oxct1 expression is regulated during transcriptional regulation and SCOT activity is regulated by post-translational modification. Their modulators and signaling pathways are schematically shown in Oxct1 gene expression in normal liver was inhibited by microRNA 122 during fetal-to-neonatal transition and trimethylation of histone H3 at lysine 27 (H3K27me3) [Oxct1 mRNA and SCOT protein expression in the heart and muscle of animals, presumably via PPARα [Oxct1 mRNA and SCOT protein expression was decreased in the failing heart myocardium and cardiomyocytes by oxidative stress [In the case of ketolysis, the main regulatory points include expression of the SCOT gene and activation of the SCOT protein, a key enzyme that catalyzes the rate-limiting step . Two isond Oxct2 . It was etolysis ,63. The  tissues . Like HM3K27me3) ,64,65. T3K27me3) . Persistia PPARα . It was −) may be responsible for nitration modifications of SCOT proteins [The SCOT protein was found to be hyperacetylated in the brain of SIRT3 knockout mice, which correlated with decreased ketolytic capacity ,67. SCOTproteins ,71.The main function of KBs is to provide extrahepatic tissues with the energy they need more efficiently during nutrient deprivation ,73. In pKBs exert many beneficial effects, such as protection against renal and hepatic ischemia/reperfusion injury, chronic inflammation-induced cardiovascular diseases, and diabetic retinal damage, by functioning as anti-inflammatory signaling molecules ,79,80,81HCAR2 is a Gi/o type of cell surface G-protein coupled receptors (GPCRs) and is abundantly expressed in various cells, including macrophages, monocytes, dendritic cells, adipocytes, and epithelial cells ,85. The Although BHB was associated with anti-inflammatory effects in most studies ,89,92,94+ memory T cells [KBs, including BHB, exert beneficial effects in part by regulating gene expression through epigenetic and post-translational modifications of histones and nonhistone proteins ,100. Som T cells . Interestingly, nonhistone proteins also undergo β-hydroxybutyrylation ,107 and An important regulatory function of KBs is its effect on reactive oxygen species (ROS) metabolism and maintenance of redox homeostasis. The antioxidant effects of KBs have been widely studied both in vitro and in vivo ,112,113.BHB can act as a direct antioxidant against hydroxy radicals . BHB canMetabolic reprograming of cancer cells is associated with their rapid growth and proliferation ,28,29. TDuring glucose starvation, normal cells use KBs as metabolic fuel. It has been shown that many tumor cells are inefficient at utilizing KBs due to dysfunction of mitochondria and/or deficient ketolytic enzymes A 44,124,126. It HMGCS2 transcription and its protein expression in c-Myc-dependent colon and rectal tumors. Consistently, HMGCS2 protein expression was downregulated in colorectal cancer patients [HMGCS2 gene by DNA hypermethylation has been shown to reduce ketogenesis and facilitate tumor cell motility in clear cell renal cell carcinoma (ccRCC) [HMGCS2 mRNA in HCC [The production of KBs by cancer cells has been shown to inhibit or promote the growth and proliferation of cancer cells. Therefore, questions about the importance of ketogenesis in cancer cells have been raised. Here, we review how key enzymes/proteins involved in KB metabolism and signaling pathways are altered in cancer cells. Interestingly, the expression and activity of HMGCS2 was shown to decrease in cancer cells, resulting in low BHB production B. Exprespatients . Similarpatients ,133,134. (ccRCC) . Similar (ccRCC) . BecauseA in HCC . HMGCS2 HMGCS2, ACAT1, HMGCL, and BDH are suggested to be potential tumor suppressors.Similarly, it was shown that other ketogenic enzymes such as ACAT1, HMGCL and/or BDH1/2 levels were significantly downregulated in ccRCC cells and patients , acute mExpression of HCAR2 has shown to be suppressed in colon and mammary tumors due to epigenetic mechanisms involving DNA hypermethylation ,138. TarIn contrast, it was reported that HMGCL expression was upregulated in oncogenic mutant BRAF V600E-expressing cancer cells and the increased AcAc promoted activation of mitogen-activated protein kinase kinase 1 (MEK1)-extracellular signal-regulated kinase 1/2 (ERK1/2) signaling that drives tumor cell proliferation and growth . A recenTaken together, dysregulated ketone body metabolism and its significance in cancer cells may vary depending on the cell type, grade and stage of the tumor, and the genetic mutation. Since cancer cells prefer glycolysis and use more glucose than normal cells, diets targeted to the Warburg effect can starve cancer cells by reducing blood glucose levels ,145,146.KDs lower blood glucose and provide KBs to tissues, thereby reducing cancer cell proliferation, and enhancing survival. KD alone or in combination with CR has been successful in the treatment of malignant gliomas in animal models ,152,158 Although a KD provides anti-tumor benefits on its own for certain types of cancer, it is much more effective when combined with chemotherapy and radiation ,173,174.In contrast, a KD showed the protumor effect in other studies including kidney cancer in a rat model of tuberous sclerosis . It was However, a recent study showed that in mice bearing BRAF V600E mutant and NRAS Q61R mutant xenografts, KBs had no direct effect on melanoma cell growth, whereas a KD slowed tumor . This suCollectively, extensive studies support the beneficial effects of KDs in reducing tumor growth, preventing cancer, enhancing the efficacy of cancer therapies, reducing the side effects of cytotoxic treatments, and increasing survival ,145,148.KBs and a KD appear to show therapeutic potential in cancer treatment ,184, but+ colon cancer mouse model [Potential mechanisms for KBs and KDs in cancer therapy are schematically shown in se model ,187. BHBse model .+ colon cancer model [KBs and/or KDs have been shown to increase ROS levels in cancer cells. BHB and KD reduced cell proliferation and induced apoptosis through ROS production in glioma-stem-like cells  and CT26er model . Therefoer model ,188. Hower model . CR and er model . BHB mayer model ,33.Administration of KDs in mouse models has been shown to enhance the anti-angiogenic efficacy of cancer therapies ,190. KDsInflammation is associated with tumor progression and resistance to cancer treatment . Cancer + T cells in malignant glioma mouse model and reduced the expression of T cell co-inhibitory receptors CTLA-4 and PD-1 and their ligands CD86 and PD-L1 [KDs and KBs have been shown to affect the immune system and reverse tumor-mediated immune suppression in malignant tumors ,193. KDsnd PD-L1 . These snd PD-L1 . AMPK isFinally, in addition to anti-tumor effects, KBs have shown to suppress protein catabolism during starvation and LPS-stimulated inflammatory condition ,195. TheThe expression and activity of key rate-limiting enzymes for ketone body metabolism are regulated by transcriptional regulation and post-translational modifications. In addition to their role as metabolic fuels in extrahepatic tissues, KBs can exert many other beneficial effects such as neuroprotection, cardioprotection, anti-aging, and anti-cancer. KBs serve as signaling molecules regulating inflammation and immune function through receptor-dependent and independent pathways. KBs also exert beneficial effects by inhibiting ROS production and regulating gene expression and other cellular functions through post-translational modifications of histone and nonhistone proteins. Through these effects, KBs have a high therapeutic potential for various diseases including cancer.Cancer cells reprogram glucose metabolism, such as the Warburg effect, for rapid cell proliferation and tumor growth. KB metabolism is also dysregulated in many types of cancer. KDs may exert anti-tumor effects through a variety of mechanisms, including elevated KBs and decreased blood glucose. Data from preclinical and clinical trials have shown strong support for the use of KDs in preventive and adjuvant cancer therapy. However, the molecular mechanism involved in the regulation of cancer biology by KBs is not fully understood. Further studies are needed to clarify the roles and mechanisms of KBs and KDs in various cancer types and cancer treatment regimens.


It plays a very critical role in many pathophysiological processes in the body. It can be endogenously produced through many enzymes analogous to the cysteine family, while the exogenous source may involve inorganic sulfide salts. H2S has recently been well investigated with regard to the onset of various carcinogenic diseases such as lung, breast, ovaries, colon cancer, and neurodegenerative disorders. H2S is considered an oncogenic gas, and a potential therapeutic target for treating and diagnosing cancers, due to its role in mediating the development of tumorigenesis. Here in this review, an in-detail up-to-date explanation of the potential role of H2S in different malignancies has been reported. The study summarizes the synthesis of H2S, its roles, signaling routes, expressions, and H2S release in various malignancies. Considering the critical importance of this active biological molecule, we believe this review in this esteemed journal will highlight the oncogenic role of H2S in the scientific community.Hydrogen sulfide (H H2S plays an essential role in mediating the many molecular, physiological, and pathophysiological functions in living systems [2S is considered to be involved in developing and progressing various diseases, ranging from non-malignant [Like nitrogen oxide (NO) and carbon monoxide (CO), hydrogen sulfide  injury [2S in cancer growth and development. In recent years, several reports have recommended that the endogenous or exogenous production of H2S may establish two different roles in forming cancer cells [Previous studies have shown that H) injury ,34,35, v) injury ,37, anti) injury ,39, anti) injury , and car) injury . Howeverer cells ,43. 2S from cysteine and homocysteine [2S may be oxidized by sulfhydryl reductase (SQR) and may be methylated through sulfhydryl-S-methyl transferase in the cytoplasm [2S is emitted by biological liquids when it joins molecules with metal and methemoglobin [2S production increased in vivo in the presence of phosphodiesterase inhibitors, inorganic sulfide salts, and organic H2S donors [2S donors [2S releasing moiety) [2S [2S in altered cells and the events that lead to its variation in tumor cells, existing research proposes that only a small amount of H2S is required for maintaining cellular activities, and that any change in its level, whether increasing or decreasing, has a significant impact on cancer-modulating cellular activities.According to Chiku et al.’s study, human CSE profligacy occurs in various reactions that produce Hcysteine . In the cysteine . In mitoytoplasm ,47. Addimoglobin . H2S proS donors . Sodium S donors ; SG-1002S donors , NOSH-as moiety) ,53, and ety) [2S ,54. Alth2S enzymatically [2S from L-cysteine. Both enzymes are co-localized in the cytosol and mitochondria [2S in both brain and kidneys [2S is instantly released or converted to acid-labile sulfur or bound sulfane sulfur and stored in mammalian cells [2S can occur through mitochondrial oxidation to sulfate and thiosulfate, excretion from the kidneys or lungs, hemoglobin-mediated scavenging, and thiol methyltransferase and rhodanese-mediated methylation to generate methanethiol and dimethyl sulfide [Recent experimental research has shown that all mammals, including humans, produce Hatically . The comchondria ,57,58. A kidneys   or phosphogluconate through nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (<10%) [2S—are formed. The direct reduction in glutathione and elemental sulfur also produces H2S. Elemental sulfur is reduced to H2S by reducing equivalents of the glucose oxidation pathway such as NADH or NADPH [2S formation in a thiol-dependent manner; for example, diallyl trisulfide (DATS), diallyl disulfide (DADS), S-allyl cysteine (SAC), and diallyl sulfide (DAS), respectively [2S is produced [Non-enzymatically, He (<10%) ,63. Whenor NADPH ,66. Furtectively ,63,67,68produced ,69 , but also in plasma. Furthermore, in germ-free mice compared to conventional animals, the traditional animals showed higher levels of bound sulfane sulfur in the fat, lungs, and plasma, with lower cysteine levels and higher CSE activity in most tissue organs [2S, which may be produced or generated endogenously, or from nutritional intake [2S endogenously are CBS, CSE, and 3-MST [The production and biogenesis of H tissues . Specifie organs . Additiol intake . The maind 3-MST .2S-producing enzymes in cancer has been studied comprehensively, and it has been revealed that cancer cells upregulate the ability of H2S-producing enzymes to assist with bioenergetics functions, in order to exploit ATP generation for the purpose of increased growth, migration, and proliferation [Different studies have revealed the altered expression and role of these three enzymes in developing several cancer cells, as explained below and summarized in feration ,83,84,852S-producing enzymes, CSE and 3-MST, showed no upregulation in tumor cells. With the consequent testing of HCT-116, HT-29, and Lovo (colon adenocarcinoma-derived cell lines), it was observed that CBS is selectively upregulated, in concert with NCM356 . Cell fractionation studies were also conducted to investigate CBS location in colon cancer cells, with the results showing that CBS is habitually localized to the cytosol, but mitochondrial translocation is also possible [2S. This response was inhibited using the prototypical CBS inhibitor compound amino-oxy-acetic acid (AOAA) [In 2013, a study comparing human colon cancer specimens with healthy mucosa tissue presented selective up-regulation of CBS in cancer tissue, whereas low CBS expression levels were observed in non-cancerous peri-tumor tissue. The other Hpossible ,87. A std (AOAA) .CBS was overexpressed in primary epithelial tissues, ovarian cancer, and multiple ovarian cancer cell line specimens similar to colon cancer cells. After examining 200 primary epithelial ovarian cancer patients’ tissues using microarrays, Bhattacharyya and colleagues found high levels of CBS expression, predominantly in a common histological variant of serous carcinoma. Tumors with higher-grade cancers and serous histology contain higher levels of CBS. Previous studies have shown strong levels of CBS expression in the FIGO stages (I and II) of ovarian cancer . In furt2S-producing enzymes for multiple cancers. Guo et al. reported that CBS and CSE could be found in prostatic epithelium normal tissue, while only CSE was found per acinar stroma cells. An androgen-dependent prostate cancer cell line (LNCaP) demonstrated noticeable degrees of CSE and CBS expression.In melanoma cancer cells that express more CBS than usual, Panza and colleagues identified the various types of congenital nevi . According to a study of primitive human melanoma, CBS and 3-MST were also present at a high but variable level. In contrast, different human melanoma cell lines  with normal epidermal melanocytes (NHEM) do not exhibit CBS expression . Despite2S synthesis in the cancer cell lines was not investigated.In contrast, an epithelial cell line of the normal prostatic peripheral zone (RWPE-1) showed low observable expression of both CBS and CSE . Similar2S in tumor angiogenesis. Unlike CBS, suppressing CSE (whose expression was intact in colon cancer) did not affect tumor development or bioenergetics. In conclusion, H2S produced from CBS serves to (i) maintain colon cancer cellular bioenergetics, thereby supporting tumor growth and proliferation, and (ii) promote angiogenesis and vasorelaxation, consequently providing the tumor with blood and nutrients. CBS-derived H2S has been identified as a tumor growth factor and anticancer medication target [CBS knockdown of cancer cells inhibited xenograft development and neovessel density, indicating a function for endogenous Hn target . CBS detn target .2S development [Like non-malignant colon mucosa cells, epithelial cell lines from colon cancer exhibit selective CBS upregulation and increased Helopment . The expelopment ,94. In aelopment . Recent elopment . Furtherelopment . CBS is elopment .CBS expression can also contribute to the development and progression of human glioblastomas . CBS exp2S suppressed by AOAA, a prototypical CBS inhibitor [Szabo et al. compared human colon cancers with normal tissue from the patient’s lining. Further, several cell lines derived from the colon’s adenocarcinoma were examined , includinhibitor . Szabo enhibitor .2S proliferative and pro-migratory characteristics are likely suitable for Akt/PI3K signaling stimulation, as early studies have established that H2S exogenous donors excite the migration of HCT116 cells via pathways activation [2S produced by CBS results from its stimulation of mitochondrial and bioenergetic actions. Additionally, Szabo et al. revealed that silencing CBS and inhibiting AOAA decreased the bioenergetic activity of HCT116, which requires basal electron transport. The respiratory reserve capacity bioenergetic parameter is an increase in mitochondrial oxygen consumption in response to a mitochondrial uncoupling agent [2S’s documented stimulation effect on GAPDH activity [Moreover, S-Adenylyl-L-methionine (SAM), an allosteric CBS activator of this compound, enhances the proliferation of HCT116 cells at low concentrations ,86. By ctivation . In summng agent . CBS inhng agent , a findiactivity , a key eExperiments with nude xenografts of HCT116 cells or patient tumor tissue (PDTX) confirmed the results in vivo. A combination of the pharmacological inhibition of CBS with AOAA and CBS silencing significantly reduced the formation of tumor xenografts. Szabo et al. have reported CBS inhibition effects in vivo . In some2S in inhibiting the proliferation, migration, and invasion of ovarian cancer in vitro, using a combination of genetic and pharmacological approaches. The regulation or inhibition of CBS has been shown to inhibit cell proliferation, and treatment with AOAA, particularly at higher concentrations, also reduces cell viability. As Bhattacharyya and colleagues investigated the intracellular mechanisms underlying these acts, they discovered that controlling or inhibiting CBS reduces the essential antioxidant glutathione’s (GSH + GSSG) intracellular content and causes apoptotic cascades. The absorption of the intracellular antioxidant may cause this latter effect after CBS inhibition/suppression. Another inevitable result of CBS cessation or inhibition is an increase in reactive oxygen species in the cells; this may be secondary to antioxidant depletion or correlated with mitochondrial function changes. Finally, CBS styling impacts on intracellular signaling in A2780 cells: Bhattacharyya and colleagues discovered that CBS suppression increased the expression of p53 while reducing the expression of the NF-kB RelA/p65 subunit [The expression of CBS is already identifiable in most of the early stages (FIGO Stage I and II) of the ovarian cancers studied . Another subunit .2S increases mitochondrial function and cell bioenergy in ovarian cancer cells, as seen in colon cancer cells. According to Bhattacharya and colleagues, CBS reduced oxygen consumption in mitochondria, and similar effects were observed when ovarian cancer cell lines were treated with a CBS, AOAA inhibitor. In addition to CBS suppression, CBS inhibition has been shown to increase mitochondrial ROS production, decrease NAD/NADH ratios, decrease ATP synthesis, and increase ADP/ATP ratios [HP ratios . The resP ratios . BhattacP ratios .2S/CSE is also involved in hepatocytes, as they correlate with H2S output and are critical for cell proliferation [2S [CSE has been shown to play a critical role in various types of cancer cells, as indicated by a study inhibiting CSE by shRNA, or a study inhibiting DL propargyl glycine and cancer cell proliferation and invasion in the human colon SW480 . Human cferation . CSE’s eferation . A CSE etion [2S . Molecultion [2S .2S in larger amounts than other colon cancer cell lines, which is consistent with the conclusion that H2S promotes cell proliferation.The expression of CSE and CBS is also reduced in prostatic cancer cell lines . The cyt2S-producing enzyme in peripheral tissues, based on their studies of mice lacking CSE [2S pathways in prostate cancer. There have been reports of CSE-induced outcomes in melanoma [2S donors foremost to cell apoptosis in both types of cancer. However, it is still unclear whether CSE enhances or prevents cancer. A recent study did not explain what CSE inhibition or removal means in these cells. Studies have shown that CSE helps cells to survive in hepatocellular carcinomas [Researchers have indicated that CSE may be the main Hking CSE . CSE hasking CSE , late-onking CSE . The oveking CSE . This sumelanoma  and gastmelanoma ,111, andrcinomas ,112. Yinrcinomas . Severalrcinomas . It alsorcinomas .NaHS stimulated cancer cell proliferation by activating extracellular signal-regulated kinase pathways in HCT116 and SW480 . In rece2S, rather than CSE [2S than controls [3-MST is expressed by all somatic cells. The expression of 3-MST by tumor cells is not unexpected. Human neoplastic cell lines have also been reported to express and activate 3-MST, with activities that are higher than CSE. Therefore, it is presumed that 3-MST is the primary source of Hthan CSE ,115. Addthan CSE ,117, glithan CSE ,34,118, than CSE , urothelthan CSE ,120, lunthan CSE , and melthan CSE  with marthan CSE . Accordicontrols . A relatcontrols . The expcontrols ,120, colcontrols , oral sqcontrols , lung cacontrols , and adecontrols  was sign2S in reaction from 3-mercaptopyruvate, were prominently suppressed [In 1960, Kun’s group reported an initial investigation using the metabolic reaction of 3-mercaptopyruvate in tumors and healthy cells. During their investigations, the author claimed that the 3-MST system in cancer cells appears to have reduced enzymatic activity than normal tissues . Later oppressed . The stippressed . An addippressed .2S by endothelial cells has been shown to occur during cell proliferation, migration, vascular relaxation, and angiogenesis, specifically under hypoxic conditions [Instead of reducing 3-MST, silencing enhanced the bio-energetic function of these cells. Due to the effects of 3-MST silencing, the bio-energetic variations may change in a specific direction. There may also be differences in cell types . Interesnditions . Howevernditions .2S. Studies with 3-MST inhibitors and 3MT silencing should be anticipated in terms of its effects on the redox processes that 3-MST regulates, in combination with the H2S effects that 3-MST induces. Simulating 3-MST would impair interactions between 3-MST-mediated proteins, possibly resulting in functional consequences. 3-MST is physically associated with enzymes such as L-cysteine de-sulfurase NFS1 and the rhodanese-like protein MOCS3 [Furthermore, experiments will be required to determine the value of these findings and the function of the tumor microenvironment. For a better understanding of tumor growth and angiogenesis, more information is needed on the possible role of 3-MST in in vivo studies . Recentlin MOCS3 .2S depends on the donor’s supplementation, cancer types, and concentration. In cancer, the function of H2S as the bioenergetics stimulator at low concentrations. According to Goubern et al., sulfides are have a high affinity to mitochondria in mammals, making them a suitable energetic substrate, even at low concentrations [2S-mediated mitochondrial respiration can only aid cancer development in the presence of sufficient oxygenation. Additionally, H2S, as a substrate for mitochondrial respiration, increases intramitochondrial cAMP levels and induces the persulfidation of mitochondrial ATP synthase and lactate dehydrogenase [2S [2S. H2S mediates persulfidation by stimulating the KATP channel and influencing downstream effects, as demonstrated by Mustafa et al. [2 promotes ischemia-induced angiogenesis in several experiments by upregulating HIF-1a expression [2S contributes to hypoxia-induced angiogenesis during cancer development is highly plausible, although insufficient evidence supports it. The pro-angiogenic activity of H2S was first discovered in the late 2000s. H2S from NaHS, when used experimentally, promotes the proliferation of endothelial cells, migration, and tubular formation [2S have also been established in rat models [2S as a physiological angiogenic agent. The PI3K/AKT pathway, mitogen-activated protein kinase (MAPK), and ATP-sensitive potassium (KATP) channels have all been shown to mediate the proangiogenic effects of H2S [2S has also been shown to promote angiogenesis in endothelial tumors. Using a well-established model of tumor angiogenesis, Puppo et al. discovered that NaHS enhances the migration of endothelial cells isolated from breast carcinomas (B-TECs). In the absence of VEGF, the inhibition of CSE inhibits the migration of B-TECs, suggesting H2S is a crucial contributor to both exogenous and endogenous breast cancer angiogenesis. In rat models of colon cancer [2S can promote tumor growth by promoting angiogenesis and by transporting nutrients and oxygen to cancer cells. However, in cancer biology, it is essential to note that high concentrations or doses of NaHS may also suppress angiogenesis [2S concentrations. H2S has an anti-apoptotic effect, including a defensive effect against apoptotic stimuli [2S-linked persulfidation.Mammal cells are currently envisaged to respond to Htrations . Thus, ttrations  for varitrations . Consequrogenase ,118. Glinase [2S , which ia et al. . H2 prompression . The hypormation . NaHS alt models . The inht models , implicas of H2S ,134. H2Sn cancer , CBS silogenesis . This in stimuli ,139. Add stimuli , hepatoc stimuli , and neu stimuli . Additio stimuli , the act stimuli , and the stimuli , mediate2S effectively and significantly reduced the expression of p21 proteins, which are considered CDK inhibitory proteins [2S acts as a proliferative factor in cancer by encouraging cell cycle progression.The cell cycle is a series of highly well-defined events that control the transition from cellular quiescence (GO) to proliferation, and attest to the high loyalty of the genomic transcript. In eukaryotes, the cell cycle is divided into four phases: gap phase 1 (G1), DNA synthesis phase (S phase), and gap phase 2 (G2), in which the cell prepares for division and the mitosis phase (M phase). Distinct chromosome separation and cell division occur during the M process . The breproteins . Similarproteins . These s2S can accelerate or prolong the cell cycle in diverse cell types, including cardiomyocytes, cancer cells, and endothelial cells [2S inhibits the expression of regulatory genes in the cell cycle and increases proliferating nuclear antigens and cyclin-dependent kinase 4 [2S was investigated for its acceleration effect on the cell cycle in colon cancer [2S on squamous cell carcinomas and colon cancer cell lines [2S-induced ERK activation. However, the molecular mechanisms of the critical role of AKT in the development of human cancer are still unclear [Cancer cells can proliferate indefinitely by evading arrest in the cell cycle. Recent evidence shows that Hal cells ,148. Herkinase 4 . H2S wasn cancer  and hepan cancer . This fun cancer ,104,105,ll lines ,104. Alt unclear . An expl2S concentrations will lead to their death, while normal fibroblasts are unaffected. 2S. H2S regulation determines the normal functioning of the cells, and depending on the cell type, any dysregulation (upregulation and downregulation) in endogenous H2S levels is associated with the development and metastasis of cancer [2S-synthesizing enzymes are increased in various human malignancies, including colon cancer, prostate cancer, breast cancer, urothelial, ovarian, oral squamous cells, and thyroid cancers, and a worse prognosis mediates the tumor to advance stages [2S levels and cancer progression, many novel types of research have recently been published in reputed journals [2S inhibition and H2S donation, respectively, on cancer cells [2S donor for breast cancer cells. HA-ADT suppresses the growth of human breast cancer cells by inhibiting the PI3K/AKT/mTOR and RAS/RAF/MEK/ERK signaling pathways [The prolonged exposure of cancer cells to high Hf cancer ,151,152.e stages ,153,154.journals ,159,160.er cells . Exposurpathways .2S synthesis out of three essential enzymes; its expression heals cancer by reversing acquired resistance to 5-FU in colon cancer cell lines [2S in cancer cells and providing a baseline with implications on the prospect of developing cancer therapies by targeting H2S levels in the human body. In adherence to the results of increased expression of Bax and Bcl-2, which mediate apoptosis-related cancer cell death and the destruction of signaling pathways upon the exposure of H2S donors achieved in [2S donor) and Dex (dexamethasone) significantly inhibits sphere formation, colony formation, and the proliferation of multiple myeloma cells by inducing apoptosis and cell cycle arrest. In addition, treatment mediated an increase in the expression of miR-127-3p and inhibited PI3K, p-AKT, and p-mTOR pathways [2S role against cancer, a novel study published on the pharmacological inhibition of H2S-producing enzymes, showed that the induction of significant changes in gene and protein expression has the potential to pharmacologically induce the mesenchymal-to-epithelia transition (MET) and disturb the EMT/MET balance in colon cancer, which evaluates and designates H2S as a potential contributor in anti-metastatic mechanisms against cancer [Similarly, promising results were achieved by inhibiting the CBS, a significant contributor to Hieved in , in recepathways . In anott cancer .While act cancer . In follt cancer , it has t cancer .2S catabolism to H2SO4 must not be abandoned entirely, as it can also lead to subsequent intracellular acidification. Consequently, uncontrollable intracellular acidification occurs in a panel of cancer cell lines, leading to cell death [2S alone was responsible for its behavior.The metabolic process of glycolysis is found within cancer cells , and it ll death . GYY4137ll death ,166, whiIn mouse models, GYY4137 has significant antitumor activity. In their study, researchers studied the impact of GYY 4137 on non-cancerous fibroblast cells such as Wi-38 and MCF10A . They di2S suppressive effect on the cell cycle switch. Sproargyl-cysteine (SPRC) acts as an H2S donor, causing G1/S phase cell cycle arrest and subsequent apoptosis in vitro and in vivo in the gastric cancer cell line SGC-7901 [2S induces cell cycle arrest in cancer cells, since neither NaHS nor GYY4137 induces cell cycle arrest in normal fibroblast cells in the above studies [Cell cycle dysregulation proved to be involved in cancer progression . Thus, cSGC-7901 . In a grSGC-7901 . FurtherSGC-7901 ,171. ForSGC-7901 . H2S ind studies ,170.2S suppressive effect is crucial for cell cycle transition, H2S donors and Sproargyl-cysteine both cause cell cycle arrest at the G1/S step and subsequent apoptosis in the gastric cancer cell line SGC-7901, in vitro and in vivo [2S tends to precisely arrest the cell cycle in cancer cells, as NaHS or GYY4137 do in normal fibroblast cells [2S causes those effects remain unknown. In eukaryotes, the cell cycle is divided primarily into three phases: G1 to S, G2 to M, and M to G1 [Dysregulation of the cell cycle has been shown to play a crucial role in cancer progression . Therefo in vivo . NaHS-in in vivo . However in vivo ,171. Lu  in vivo . To our st cells ,170. How M to G1 . The pre M to G1 . SPRC tr M to G1 . The adm2S appears to inhibit the proliferative activity of cancer cells by specifically blocking the cell cycle and protecting non-cancer cells from death [2S’s anticancer activity. For example, H2S can increase E-cadherin levels, which have anti-metastatic effects [2S on biological processes remain unknown, because H2S is responsible for a plethora of biological processes. Cancer cells disturb the balance between apoptosis and survival by activating pro-survival pathways in persistently growing cancer cells [2S has also been shown to inhibit its activation by TNF and lipopolysaccharide [2S [Several studies have found that NaHS can arrest the cell cycle and promote the expression of the p21Cip1 protein in colon cells treated for 12 to 24 h . Accordiom death . There c effects , and inh effects . The moler cells . NF-κB, ccharide ,180. Thuride [2S  or donatride [2S  causes d2S inhibits NF-κB activity is not well understood, and further research is needed to better understand the mechanism of H2S anticancer action. For instance, GYY4137 induces apoptosis in hepatocellular carcinoma cell lines by inhibiting STAT3 activators and downregulating B cell lymphoma 2 through STAT3 [2S exposure causes apoptosis in oral cancer cells, probably due to the downregulation of pleckstrin homology-like Domain-A1, an apoptotic suppressor found in this type of cancer [2S-target proteins involved in cell survival pathways must be addressed in the future.In contrast, the molecular mechanism by which Hgh STAT3 . Additiof cancer . The imp2S production and programmed cell death have been well-investigated in many studies. H2S prevents apoptosis in colon cancer cells induced by β-phenyl ethyl isothiocyanate [2S signaling pathway is crucial to maintaining the proliferation of hepatoma cells. While the inhibition of these pathways prevents these cells from developing, this may be due to mitochondrial apoptosis. However, treatment with NaHS increases cell viability in the hepatic cells, PLC/PRF/5 [2S is necessary for increasing the apoptotic ratio of cancer cells (CA9-22), apoptotic markers in normal keratinocytes are unknown [2S sulfhydrates the NF-κB p65 subunit, facilitating its attachment to the co-activator ribosomal protein S3. The anti-apoptotic capabilities of NF-κB are drastically diminished in CSE mutant mice. H2S that is released via CSE improves DNA binding and NF-κB gene activation, both being abolished in CSE-deficient animals. H2S sulfhydrates the NF-κB p65 subunit, facilitating its attachment to the co-activator ribosomal protein S3. The anti-apoptotic capabilities of NF-κB are drastically diminished in CSE mutant mice [2S appears to mediate anti-apoptosis in the progression of diseases that are associated with extreme cell development and division, including cancer. However, the exact mechanism is unknown.Programmed cell death  plays a fundamental role in controlling oncogene initiation, unrestrained proliferation, and chemotherapy. Recently, Hocyanate . 6-hydroocyanate . It has LC/PRF/5 . AdditioLC/PRF/5 . Althoug unknown . H2S sulant mice ,122. Due2S acts as a signal molecule in various structures and tissues, including the circulation, nervous system, and organs [2S in inflammatory cells [2S promotes the proliferation and migration of SW480 cells derived from human colon cancer in vitro, and can contribute to SIRT1 upregulation. CBS increased H2S synthesis in ovarian and colorectal cancers, which are critical for the bioenergetics, proliferation, and migration of cancer cells [2S increases the expression of IL-6 and IL-8 in periodontal fibroblasts, which can contribute to the stimulation and production of periodontitis [Hd organs ,118. Endry cells ,183. Honer cells . By actidontitis .2S generation. CSE enhanced p65 DNA binding and downstream gene expression in mice lacking CSE. NF-anti-apoptotic B’s activity is dramatically diminished in mice lacking CSE [2S has been proposed as an endogenous mediator of inflammation via its increase in the activity of the NF-κB pathway [2S can suppress the activation of the NF-κB pathway in inflammatory conditions [2S acts as an endogenous stimulator for the Keap1pNrf2 pathways [2S assists with DNA repair by triggering the MEK1/REK/PARP1 pathway [TNF functions as an activator of NF-κB pathways, resulting in increased CSE expression and Hking CSE . As a re pathway . Similar pathway . Howevernditions ,123,124.pathways . Activatpathways ,184. Zha pathway .−3 M NaHS, a donor of H2S for 24 h, inhibits the migration, proliferation, and division of human hepatocellular carcinoma cells, inducing cell autophagy [2S activates the AMPK-activated protein kinase (AMPK) in rat glomerular epithelial cells, BV2 mouse microglial cells, and C2C12 mouse skeletal muscle cells via calmodulin kinase beta (CamKK) [By inhibiting the signaling pathway PI3K/AKT/mTOR, treatment with 10utophagy . A recenutophagy . Numerou (CamKK) ,130. Thi (CamKK) . Excessi (CamKK) . The fin (CamKK) . Accordi (CamKK) . NaHS (0 (CamKK) .Protein sulfhydration is a post-translational protein alteration in which a sulfur atom is added to a reactive protein cysteine, resulting in a -SSH or a persulfide group being created. Cysteine persulfide is formed when an oxidized cysteine derivative combines with a sulfide or sulfide oxidation product. Protein sulfhydration predominantly inhibits, with most activating events being driven by the persulfidation-induced inhibition of a negative regulator . This seSimilarly, polysulfides are also considered to be the key players in mediating the different oncogenesis pathways. A recent study indicates that a high CBS and CSE expression level indicates a poor prognosis ,189. A sNF-kB is a dimeric transcription factor family that is triggered through a wide range of stimuli and is involved in immunological responses, inflammation, and cancer . A modif2S can operate as an endogenous activator of the Keap/Nrf2 pathway. By stimulating the Keap1-Nrf2 pathway, H2S can stimulate the expression of enzymes such as glutathione S transferases, epoxide hydrolase, and ferritin, allowing cancer cells to adapt to prolonged oxidative stress and progress [2S can aid in DNA repair, as in Zhao et al. [2S persulfates MEK1, in particular, in cysteine-341, and thereby, it influences downstream effects. Many cell types are affected by H2S, including cardiomyocytes, cancer cells, and endothelial cells [2S  lowers the expression of cell cycle control genes, such as replication protein A70 and retinoblastoma protein 1. However, in certain oral cancer cell lines, it increases the production of proliferating nuclear antigen and cyclin-dependent kinase 4, which leads to cell proliferation [2S can speed up the cell cycle in colon cancer cells [2S-induced acceleration of the cell cycle [2S-induced ERK activation. The molecular mechanism by which H2S phosphorylates AKT is, however, unknown. In light of AKT’s crucial role in creating human cancer [The NF-kB pathway inhibits apoptosis by increasing the production of anti-apoptotic proteins such as TNFR-associated factor (TRAF)-1, TRAF-2, caspase-8-c-FLP, and cellular inhibitors of apoptosis , signifyprogress ,195. Eveo et al. . H2S peral cells ,196. Forferation . Studieser cells  and hepaer cells . Cell lier cells ,104,105,ll cycle ,104. Altn cancer , deciphe2S levels, indicating a function for H2S in DNA repair. Low cellular H2S levels enhanced ATR kinase activity, as determined through CHK1 phosphorylation; high amounts of H2S, on the other hand, prevented ATR ser-435 phosphorylation, which is a hallmark of ATR kinase activity [2S concentrations, increases in ATR and CHK1 may increase H2S production. Furthermore, targeted ATR inhibition is being investigated in cancer treatment [2S production [ATR kinase suppression reduced cellular Hactivity ,197. ATRactivity ,199. Furactivity ,201. Becreatment . This caoduction . These p2S has complex and robust immune system-regulating effects in normal and pathologic conditions, with decreased functioning being frequently observed at low and high H2S concentrations [2S-induced immune modulation has a role in cancer [2S protects the cancer cells from activated macrophage-generated ROS [2S donation controls tumor development via immune system regulation [2S can stimulate and prevent tumor development by modulating the immune system.Htrations . Considen cancer . CBS, asated ROS . Melanomated ROS . MDSCs cgulation ,150. TheCell death is crucial for mammalian growth and homeostasis, and it is thoroughly interwoven with an organism’s physiological function and pathological condition . Cell de2S endogenously, acting as a cardiovascular protective enzyme and as the key enzyme for l-cysteine (a precursor of GSH) [2S has an antioxidative effect by increasing GSH content and reducing ROS [2S. Growing amounts of data suggest that H2S increases GSH synthesis to reduce oxidative damage. In a neurocyte, mitochondrial H2S synthesis raises the amount of GSH and encourages its redistribution to the mitochondria, therefore protecting the neurocyte from oxidative stress [2S increases GSH production in a myotube to ameliorate impaired glucose homeostasis [2S donor NaHS treatment boosts GSH synthesis, reducing oxidative stress and postponing cell senescence [2S generation considerably recovered GSH loss in response to HHP. As a result, H2S downregulation-mediated GSH reduction might constitute a unique mechanism of ferroptosis in HHP [2S on ferroptosis suppression have recently been demonstrated in research. H2S inhibits ferroptosis by suppressing ALOX12 acetylation and controlling the stability of the xCT , according to Wang and Chen et al. [2S donor GYY4137 reduces ferroptosis, which helps to reduce acute lung damage [2S generation with the CBS inhibitor CH004 supplement worsens ferroptosis [2S might inhibit ferroptosis, and so provide protection. The study found that administering the H2S donor NaHS boosted GPX4 expression while decreasing ROS generation and lipid peroxidation, correcting high hydrostatic-induced ferroptosis.Ferroptosis manifests in cells primarily as decreased mitochondrial volume, increased bilayer membrane density, and the reduction or disappearance of mitochondrial cristae, with no nuclear concentration or chromatin marginalization . In gene of GSH) . H2S hascing ROS . In addicing ROS ,217. GSHcing ROS . L-cystee stress . H2S inceostasis . H2S donnescence . Accordis in HHP . The benn et al. ,222. Theg damage . In heparoptosis . These i2S reduces ferroptosis, which helps to reduce HHP-induced VSMC dysfunction [2S content in colorectal cancer, thereby inhibiting the growth of CT26 and HCT116 colorectal cancer cells. Furthermore, removing H2S from colorectal cancer cells inhibits tumor growth by activating ferroptosis, a non-apoptotic form of cell death. Biosafety-related toxicological and pathological analyses demonstrated the low toxicity and high safety of VZnO in the treatment of colorectal cancer [Furthermore, NaHS reversed RLS3-induced ferroptosis. Altogether, Hfunction . Zinc oxl cancer . Traditil cancer ,227. Fer2S are being increasingly recognized. Recent studies have identified novel gas sources to restore physiological function to diseased cells or organs. To date, many individuals have been identified as donors. H2S compounds occur naturally in garlic [2S donors [2S-releasing NSAID derivatives [2S donors, including ADT-OH, NaHS, thiobenzamide, DADS, and DATS, have recently been used for indefinite endogenous H2S development [2S donor-based therapies, are being tested in phase II clinical trials [2S promotes angiogenesis, accelerates the cell cycle, prevents apoptosis, and promotes the expression of oncogenes separately [2+ buildup [The cytoprotective properties of Hn garlic ,228, suln garlic ,230. CysS donors  are cystS donors ,180,232 ivatives , and GYYivatives ,234,235.elopment ,238,239.l trials ,241. Furl trials ,246,247,parately . Tumor gparately ,250,251.parately . ITCs arparately . ITCs thparately . AITC  inhibited cell cycle development in human leukemia cells by altering chromatin histones’ acetylated and methylation states . In compFurthermore, GSTP1 methylation is crucial in tumor initiation in prostate cancer. In this context, PEITC (0.5–20 M) has been shown to reduce the deacetylation and methylation of the GSTP1 gene, hence reducing the oncogenic process . PEITC  in phase III research . In a coLenvatinib was initially characterized as a multitargeted RTK inhibitor that is capable of inhibiting several kinases at nanomole doses  of 4–100 nM in 2008 . In animThe initial phase I study, which included 27 patients , was conAnother phase I trial with 87 individuals was carried out. In a 28 day cycle, the dosage of lenvatinib was gradually increased from 0.2 mg to 32 mg once a day. Analyses of pharmacokinetics were conducted on days 1, 8, 15, 22, and 28. DLT was identified as G3 proteinuria, with an MTD of 25 mg daily . As a ren = 7) in a three-plus-three pattern, lenvatinib was increased to 18 mg (n = 11) and 24 mg (n = 2) in conjunction with 5 mg of everolimus, both given once daily. The MTD was calculated to be 18 mg of lenvatinib and 5 mg of everolimus per day [Consequently, for patients with HCC CPA 49, initial doses of 12 mg and 8 mg once daily for persons weighing 60 kg were recommended. A phase Ib study combining lenvatinib and everolimus was conducted to establish the safe dosage for RCC. Starting with 12 mg of lenvatinib once daily  H22S and associated sulfur compounds in cancer-related controls. Higher H2S, for instance, has been found in the headspace vapor of stomach contents in patients with gastroesophageal cancer. Higher H2S and methanethiol levels have been discovered in colon and lung cancer patients’ flatulence and exhaled air [2S has been used to detect cancer cells and as a cancer biomarker in mice [2S and similar sulfur compounds may be found in high concentrations in body fluids and gases, and may be helpful in cancer diagnosis. H2S and similar sulfur compounds in particular might be utilized in cancer diagnosis by measuring substances such as thiols in the blood and urine to track the efficacy of cancer treatment, induce remission, and identify recurrence.Numerous studies have detected higher levels of Haled air ,384,385.aled air . The amoaled air ,387,388.aled air . Ultimat in mice ,390. The2S in mammals. These enzymes are over-expressed in all cancer types, and show cancer-related properties. The current understanding of H2S research firmly reveals that these enzymes are a key player in regulating the proliferation, migration, and the invasion of cancer cells. In other words, CBS, CSE, and 3-MST may serve as new molecular markers and biomarkers for the diagnosis and treatment of cancer. Considering the double role of H2S in cancer, H2S donors releasing high levels of H2S and other pharmacological designed H2S inhibitors are attaining much attention, both in research stations and clinical settings. Being functionally active biomolecules, H2S has become the most highly investigated molecular target in cancer biology across the globe. As the conclusion of this review, we have stated that CSE, CBS, and 3-MST are three enzymes that play an essential role in producing H2S in important cellular events that directly and indirectly mediate cell fate, there is still much to do before using H2S-based anticancer drugs in pre-clinical trials. Firstly, no study has investigated the pathways involved in the beneficial effects of H2S donors on cancer, such as NaHS, ADT-OH, DATS, and GYY4137. Secondly, extensive research is needed on the link between the production of endogenous H2S through CBS, CSE, and 3-MST, and the activation of cyclin-dependent kinases (CDKs), as we know that CDKs have a vital role in the regulation. Thirdly, most studies are evaluating the function of H2S in conjunction with other drugs and messengers, including NO, which moderates many physiological and pathological processes. These messenger molecules offer new perspectives on cancer treatment. Thus, further research is needed to clarify its impact on different cancers.Although this review has summarized the potential involvement of H2S system (2S pharmacological inhibitors, one might assume these effects translate into functionally detectable in vivo models. Mice bearing tumors could be studied. Cancer cell signals, survival, and bioenergetics, and perhaps also angiogenesis, depend on the HS system . With th


Compared with group A, bodyweight, fat weight, ALT, TG, and hepatic steatosis were significantly increased in B and C groups (P < 0.05); compared with group B, group C showed a significant decrease in bodyweight, fat weight, ALT, TG, and hepatic steatosis (P < 0.05). Compared with group A, the expression levels of GRP78, IRE-1α, CHOP, caspase-12, and Bim in liver tissue of groups B and C mice were increased. Bcl-2 decreased (P < 0.05). Compared with group B and group C after naltrexone intervention, the expression levels of GRP78, IRE-1α, CHOP, caspase-12, and Bim decreased significantly, and Bcl-2 increased significantly (P < 0.05). Naltrexone can effectively reduce bodyweight and adipose tissue accumulation, reduce liver fat lesions, improve the expression of lipid metabolism-related proteins and endoplasmic reticulum stress, reduce liver lipid synthesis, and protect liver cells.This study aimed to explore the effect of naltrexone on the expression of lipid metabolism-related proteins in liver steatosis induced by endoplasmic reticulum stress in mice. Thirty inbred mice (C57BL/6J) were divided into three groups: group A , group B (model control), and group C . The male mice in group A were fed a regular diet, and the mice in groups B and C were fed a high-fat diet. Liver steatosis was observed by histopathological sections. Mouse liver  and triglyceride (TC)) content (glucose regulatory protein (GRP78), endoplasmic reticulum transmembrane protein kinase-1 Endoplasmic reticulum stress (ERS) will be formed once exogenous substances, hypoxia, oxidative stress, and other reactions lead to ER imbalance. Cells activate signaling pathways through the unfolded protein response (UPR), including endoplasmic reticulum transmembrane protein kinase-1tability . Hepatictability , 5. Nalt30 inbred (C57BL/6J) male mice  were placed in SPF grade animal room for one week after adaptive feeding, randomly divided into three groups, group A (control), group B (model), and group C , 10 animals in each group. The study was approved by the Affiliated Hospital of Zunyi Medical University.Naltrexone , 0.9% normal saline , high-fat feed  and conventional feed , ALT and TG test kits , BCA protein quantitative reagent box , PCR kit , xylene, absolute ethanol, and other solutions and primers , related protein corresponding antibodies , cryogenic centrifuge , nucleic acid protein analyzer , and enzyme marker .After one week of adaptive feeding, 30 inbred (C57BL/6J) male mice were randomly divided into three groups, The mice were divided into group A (control), group B (model), and group C ; each group of 10 mice were weighed and recorded once a week. The male mice in group A were fed with regular diet, and the mice in groups B and C were fed high-fat diet. Two mice were selected to establish a fatty liver model after 8 weeks of feeding. The histopathological sections were made to observe whether the model was established successfully. After the establishment of the model, a total of 24 mice, three groups of 8 mice, group A and group B were given an equal volume of 0.9% normal saline, intervention for 4 weeks, and the mice in group C were given naltrexone (100 mg/kg/d) by gavage at 8 o'clock daily according to their actual weight.Liver tissue specimens were collected; 12 hours after the treatment, the mice were fasted and fixed under anesthesia. The weight, liver weight, and fat weight of mice were measured.Blood samples were collected; 12 h after the course of fasting and under anesthesia, the mice were fixed. The abdominal cavity of mice was dissected. Cardiac blood was obtained by puncture of the diaphragm with a 5 ml syringe and placed in a 2 ml EP tube containing heparin. After 10 min, centrifugation (1500 r/min 20 min) was performed. The upper serum was placed in a low-temperature refrigerator to be tested.α (IRE-1α), C/EBP homologous protein (CHOP), cysteine-containing aspartate proteolytic enzyme 12 (caspase-12), B lymphoma-2 (Bcl-2), and cell death mediator (Bim). All operations were performed in strict accordance with the standards in the kit.Detection of mouse liver  and triglyceride (TC)) content, histopathological section (HE staining) to observe the grade of liver steatosis, and detection of lipid metabolism-related protein expression level: glucose regulatory protein (GRP78), endoplasmic reticulum transmembrane protein kinase-1t-test. The expression level of a protein associated with the lipid metabolism, ALT, and TG was expressed by (‾x ± s) (P < 0.05).Three groups of mice experimental data were included in SPSS 23.0 software analysis, and measurement data comparison is done using the P < 0.05). Compared with group B, the bodyweight and fat weight of the mice were significantly decreased after naltrexone treatment (P < 0.05), as shown in Compared with group A, the bodyweight and fat weight of group B and group C were significantly increased (P < 0.05). Compared with group B, ALT and TG were significantly decreased and the degree of hepatic steatosis was reduced in group C, as given in Compared with group A, ALT, TG, and hepatic steatosis were observed in group B and group C . Compared with group B and group C after naltrexone intervention, the expression levels of GRP78, IRE-1α, CHOP, caspase-12, and Bim decreased significantly, and Bcl-2 increased significantly (P < 0.05), as given in Compared with group A, the expression levels of GRP78, IRE-1This study shows that eating high-fat diet can cause lipid metabolism disorder and related apoptosis, but naloxone can reverse the changes of the above indicators, suggesting that naloxone may be used as a clinical treatment for abnormal lipid metabolism diseases.α gene can mediate activation of CHOP and caspase-12 and initiate ERS-mediated apoptosis. CHOP, JNK, and caspase-12 are the main apoptotic pathways mediated by endoplasmic reticulum stress. The apoptosis of hepatocytes in mice with hepatic fibrosis may be mediated by activating key signal molecules [Severe hepatic steatosis may even lead to liver failure or stem cell necrosis, so clinical attention to the treatment of hepatic steatosis is high , 7. Endoolecules , 12.P < 0.05). Compared with group A, the expression levels of GRP78, IRE-1α, CHOP, caspase-12, and Bim in liver tissue of groups B and C mice were increased. Bcl-2 decreased (P < 0.05). The reason is that after a high-fat diet, mice will enter the liver with excessive lipid and fatty acid content, cause endoplasmic reticulum stress reaction, affect the related lipid metabolism-related protein levels, and continue to aggravate hepatic steatosis [P < 0.05). Compared with group B, group C after naltrexone intervention, the expression levels of GRP78, IRE-1α, CHOP, caspase-12, and Bim decreased significantly and Bcl-2 increased significantly (P < 0.05). It can be seen that naltrexone can improve endoplasmic reticulum stress response. Liver steatosis is a kind of reversible injury, and the original nature of liver fat can be restored after naltrexone eliminates the cause of the disease. The reason may be that naltrexone can reduce the stress on the endoplasmic reticulum and inhibit the degree of apoptosis of liver cells by interfering with the main apoptosis pathways of ERS mediated by IRE-1, CHOP, and caspase-12 pathways. In addition, naltrexone can also reduce the therapeutic effect of liver fatty disease by changing the phenotype and function of macrophages and other mechanisms.The results of this study showed that compared with group A, after a high-fat diet, the bodyweight, fat weight, adipose index, ALT, and TG in groups B and C were significantly increased, and hepatic steatosis was observed (teatosis . The phateatosis –16. The In conclusion, naltrexone can effectively reduce bodyweight and adipose tissue accumulation, reduce liver fat lesions, improve the expression of lipid metabolism-related proteins and endoplasmic reticulum stress, reduce liver lipid synthesis, and protect liver cells.