Extensive research has been carried out to understand and elucidate the mechanisms of paraoxonase 1 (PON1) in the development of diseases including cancer, cardiovascular diseases, neurological diseases, and inflammatory diseases. This review focuses on the relationship between PON1 and cancer. The data suggest that PON1, oxidative stress, chronic inflammation, and cancer are closely linked. Certainly, the gene expression of PON1 will remain challenging to study. Therefore, targeting PON1, redox-sensitive pathways, and transcription factors promise prevention and therapy in the development of several diseases, including cancer. Oxidative stress is an imbalance between the production and accumulation of free radicals and reactive metabolites, generally known as “reactive oxygen species” (ROS) in cells and tissues. ROS are typically generated as by-products of oxygen metabolism, which perform several physiological roles . Despite this, xenobiotics significantly increase ROS production, causing imbalances that lead to oxidative stress . Under aElevated levels of ROS are thought to be oncogenic, damaging DNA, proteins, and lipids, and promoting genetic instability and tumorigenesis ,5. ROS aROS are also responsible for the development of several diseases, including chronic inflammation, diabetes mellitus, and atherosclerosis ,9,10. CaCancer is a disease of cellular mutation, proliferation, and aberrant cell growth, usually defined by at least three stages—initiation, promotion, and progression—that generally involve oxidative stress. The first stage of cancer (initiation) is defined as a stable, heritable change. This stage is irreversible and results in a mutational event produced by physical agents, chemicals, and ROS . AdditioConsiderable evidence has demonstrated the role of antioxidants’ protective mechanisms based on enzymatic and nonenzymatic antioxidant molecules to link chronic inflammation and cancer ,21. In hThe most efficient enzymatic antioxidants involve superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GRx) ,22,23. RInflammation is a natural mechanism of the immune response to control infection, restore damaged tissue, or respond to stress control . TraditiAs indicated in Neutrophils are the most abundant cells in innate immunity, usually recruited to infection or damaged sites by responding to chemoattracting signals . Once inHowever, macrophages are highly plastic cells that change their phenotype and respond to various environmental factors. Circulating monocytes, the precursors of macrophages, pass through the vascular endothelium to mature into macrophages in the peripheral tissues . MacrophNeutrophils and macrophages produce ROS. Mitochondria mainly produce ROS, usually cleared by antioxidants, such as glutathione, vitamins C and E, and some enzymes such as catalase and peroxidase . ROS indROS can cause oxidative damage of cellular macromolecules ,58, whicDendritic cells (DCs) are antigen-presenting cells involved in priming and activating naïve T lymphocytes. They are also essential for sensing pathogens because they express PRR which recognizes PAMP or DAMP, leading to the secretion of IL-1β, TNFα, IL-6, and IL-12 ,65.The adaptative immune cells are also contributors to the inflammatory process. T-helper cells differentiate in effector cells after receiving signals from the T cell antigen receptor (TCR), cytokines, and costimulatory signals. T-helper effector subsets include TH1, TH2, TH9, TH17, and TH22 cells . TH17 ceResolution of the inflammatory process involves macrophages, namely phagocytes, that detect the apoptotic cells and secrete IL-10  and reguAlthough acute inflammation contributes to the protective host response, chronic diseases could result from uncontrolled inflammation and failure of immune cells to restore homeostasis. The imbalance between pro- and anti-inflammatory cytokines contributes to inefficient regulation of the inflammatory process . There iHowever, tumor cells also secrete some factors that serve as the immune response evasion mechanisms to avoid apoptosis; for example, the secretion of different molecules that promote regulatory T cells recruitment, polarize macrophages to an M2 phenotype , or promThe inflammatory response is associated with genes involving balanced and regulated pro- and anti-inflammatory effects . The proNF-κB transcription factors are implicated in regulating normal cellular and organismal processes, such as immune and inflammatory responses, developmental processes, cellular growth, and apoptosis .Dysregulated NF-κB activity causes inflammation-related diseases and cancer. Two pathways activate NF-kB: the canonical pathway, which responds to an external stimulus involved in inflammation, immune response, cell proliferation, differentiation, and survival; and the second, the non-canonical pathway, associated with the development of cells in multiple layers . For actPRRs expressed in innate immune cells recognize microbial PAMPs or DAMPs, therefore inducing the expression of proinflammatory cytokines, such as TNF-α , IL-1β, IL-6, IL-8, IFN-I (type I interferons), chemokines, and anti-microbial proteins, which leads to the inflammatory response ,100,101.However, cell differentiating or developmental stimuli, such as BAFF, RNAKL, or lymphotoxin, activate the non-canonical NF-κB pathway to induce RelB/P52 dimer in the nucleus. In this pathway, ligands induce NF-κB, encouraging kinase (NIK) activation. NIK phosphorylates NF-κB2 protein leads to proteasomal processing of the NF-κB2 precursor protein p100 into the mature p52 subunit. Then, the p52 dimerizes with RelB to regulate a distinct class of genes . Some studies have suggested a role for STAT family proteins, especially STAT3, in inducing and maintaining a pro-carcinogenic inflammatory microenvironment. STAT3-mediated malignant property is associated with chronic inflammation. Cytokines, chemokines, and other mediators are essential for inducing and maintaining a cancer-promoting inflammatory environment, where STAT3 regulates their expression. Moreover, it has been suggested that NF-κB and STAT3 act as two major transcriptional factors linking inflammation with tumorigenesis and functionally interacting with each other . TherefoTNF-α. Additionally, AP-1 is related to asthmatic inflammation, cardiovascular disease, cancer, and others [NF-κB and AP-1 are essential in regulating immune genes and inflammation progression. However, it has been reported that the anti-inflammatory mechanism involves the inhibitions of a transcription factor, such as NF-κB, STAT, MAP, and AP-1. AP-1 is a heterodimeric protein composed of proteins ATF and JDP, c-Jun, c-Fos, capable of modulating the expression of genes, such as d others ,104.The hypoxia-inducible transcription factor (HIF) plays a fundamental role in metabolic changes, which conduct cellular adaptation to low oxygen accessibility through the transcription of genes possessing an HRE. Cellular HIF expression can be modulated by hypoxia, inflammation, cancer, among other factors ,106. EviCyclooxygenase is a key enzyme in synthesizing prostaglandins through the oxidation of arachidonic acid. Prostaglandins are related to inflammation, pain, the development of cancer, and some functions of homeostasis in various organs .There are two isoforms of cyclooxygenases, COX-1 and COX-2, encoded by different genes, which are also located in different cell sites with different affinities for their substrate. COX-1 is expressed constitutively and plays a major role in the different homeostatic processes. In contrast, COX-2 has a constitutive expression in organs, such as the brain, kidney and an inducible enzyme. The metabolites of COX-2 are involved in inflammation and pain induction ,108. AddCOX-2 gene contains typical regions, such as those of early response genes, which permit fast upregulation when stimuli that trigger cytokines such as TNF interleukin-1 are present. Additionally, the COX-2 mRNA has labile sites encouraging degradation of the fast product and consequently decreasing gene expression when the stimulus is withdrawn. The COX-2 upregulation gene is observed in cell types such as macrophages and endothelial cells, among others. However, glucocorticoids and other anti-inflammatory agents decrease the COX-2 gene expression [The pression ,109. Although a large diversity of elements control the expression of COX-2, two important elements are b-catenin (directly by binding to the promoter or indirectly) and NF-kB. Additionally, the COX-2 mRNA is regulated at the post-transcriptional level through translation mechanisms and stability, mediated by AU-rich sequence elements. Several proteins play a crucial role in these events; among them are AUF1, HuR, CUGBP2, hnRNPA1 TIA1, TIAR, and TTP . The data reported in the literature show the relevance of RNA-based processes in controlling inflammatory responses. Additionally, non-coding RNAs (ncRNAs) have been reported in inflammation regulation. In this regard, long non-coding RNAs can mediate the interaction between DNA and proteins or modulate the translation (through miRNAs or mRNAs) .The proinflammatory state is characterized by high circulating levels of proinflammatory markers, including IL-1β, IL-1 receptor antagonist protein (IL-1RN), IL-6, IL-8, IL-13, IL-18, CRP, IFNα, and IFNβ, transforming growth factor-β (TGFβ), TNF and its soluble receptors (TNF receptor superfamily members 1A and 1B), and serum amyloid A .Higher circulating proinflammatory cytokine levels confer deleterious effects on the remote organs. Most comorbid conditions such as hypertension, diabetes, obesity, and dyslipidemia annotated with cardiovascular risk have chronic inflammation associated with elevated levels of proinflammatory cytokines, in particular, TNFα, showing a more significant impact on cardiac function and remodeling ,115,116.PON1 is recognized as an antioxidant enzyme that inhibits LDL oxidation. Furthermore, it is accepted as an enzyme with anti-inflammatory features that limit the release of proinflammatory mediators . It is wNevertheless, the activity of this enzyme is modulated under oxidative stress conditions . FurtherApoA1 and PON1 mRNA. There is a nuclear factor-κB (NF-κB) binding site on the promoter of SAA but not in ApoA1 or PON1 promoters. The inhibition of NF-κB decreases SAA mRNA and consequently increases the ApoA1 and PON1 mRNA levels [PON1 gene at a transcriptional level through the AKT/IKK/NF-κB activation pathway in human hepatocyte-derived HepG2 cells [As mentioned above, studies have shown that hepatic PON1 is regulated in response to inflammatory cytokines. Treatment with lipopolysaccharide and oxidized LDL in animal models and human hepatocytes reduced the mRNA levels of PON1. This effect was similar in the presence of TNFα, IL-1β, and IL6 ,127. MorA levels . The resA levels ,129. In G2 cells . PON1 anG2 cells ,131. StuG2 cells ,132.Additionally, some studies in humans have shown a relationship between cytokines and low PON1 activity. For example, Kerekes et al.  observedRecently, PON1 has been suggested to protect cells from oxidation by inhibiting the synthesis of chemokine (C-C motif) ligand 2 (CCL2) and the subsequent inflammatory reaction. However, excessive production of ROS would inhibit PON1 and increase CCL2 production. This induces the migration and infiltration of immune cells in target tissues, disturbing normal metabolic function. This disruption involves pathways controlling cellular homeostasis and metabolically driven chronic inflammation, stating its relationship with the development of some diseases .2+-dependent A- esterase with a molecular mass of 43 kDa and 354 amino acids [The human PON1 is a circulating Cano acids ,136. PONno acids . This enno acids ,138; howno acids , especiano acids ,140.PON1 received its name from its ability to hydrolyze paraoxon (diethyl p-nitrophenyl phosphate), an active metabolite of the insecticide parathion ,143,144.The original function of PON1 was in lactonase (LAC), since lipophilic lactones constitute its primary substrates, shown to hydrolyze over 30 different lactones (cyclic esters) ,149,150.2O2 (via its peroxidase-like activity), preserves HDL integrity and function, and prevents LDL oxidation and the cell membrane; it is thought to be atheroprotective [Pseudomonas [Additionally, several physiological functions have been attributed to PON1, particularly its degradative capacity of lipid peroxides ,152,153,otective . Additioudomonas ,155,156.The expression and activity of PON1 have been associated with lung cancer, multiple myeloma, and papillary thyroid cancer ,158,159.PON1 gene is located between q21.3 and q22.1 on the long arm of chromosome 7 in humans (chromosome 6 in mice) [PON1 gene size is approximately 26 kb, which comprises nine exons and an extra codon at position 106 (lysine) in exon 4 [PON1 appears to be a non-canonical polyadenylation signal sequence. The fourth intron (of eight) contains a CA repeat, the length of which is polymorphic, and allele lengths vary by up to four CA units [The in mice) ,137. PONn exon 4 . PON1 apCA units .PON1 gene does not contain a canonical TATA box in the 5′ UTR but is rich in GC sequences [PON1 gene has 200 polymorphisms distributed between the promoter and its coding region, but most are not characterized. It has been described that the polymorphisms of the promoter impact the regulation and expression of the gene. However, the polymorphisms of the coding region have a more significant impact on its activity in serum. A potential role for the transcription factor specificity protein 1 (Sp1) was demonstrated by Deakin et al. [PON1 expression. However, one set of transcription factors with which also act in tandem are the sterol-regulatory-element-binding proteins (SREBPs) [The promoter sequence of the equences . Furthern et al. . Sp1 is (SREBPs) . TherefoPON1 mRNA is expressed in the liver, with the PON1 glycoprotein consisting of 354 amino acids (molecular mass of 43-KDa). It retains its hydrophobic signal sequence in the N-terminal region , which enables its association with HDL [with HDL . Furtherwith HDL . Therefowith HDL . Further2+, one essential for activity and interaction with the side chain oxygens  as an oxyanion that stabilizes the substrate and necessary for optimal hydrolytic activity. The second ion is essential for stability with its removal causing irreversible structural disruption of protein structure [An X-ray crystallography study has indicated the structure of PON1 as a 6-bladed propeller, with a lid covering the active site passage and containing two Catructure ,166. Thetructure ,168.PON1 gene has nearly 200 SNPs. The main polymorphisms identified in the promoter region of the PON1 gene are −909 (G or C), −832 (A or G), −162 (A or G), −126 (C or G), and −108 (C or T). −108C, −832A, −162A and −909G have higher levels of expression than −108T, −832G, −162G, and −909C. The literature that identified the positional changes reported these polymorphisms  due to the variations in the examined sequences. Despite the physiological relevance of the coding region polymorphisms, an association with highly significant regulatory region polymorphisms in PON activities has been described in the case of the −107C and −824A polymorphisms. The −107C and −824A variants have higher activity than the −107T and the −824G variants [PON1 transcription, whereas mithramycin acts as an inhibitor [PON1 [PON1 gene; however, their significance is not yet studied. A significant increase in serum PON1 mass and activities between haplotype subgroups could be explained by a unit increase in the number of high-expresser variants of the −108 site (−108C) alone. Several studies have revealed that the polymorphisms at positions −126 [The variants ,169. Howvariants . It is bvariants . The −10nhibitor . The preor [PON1 ,172. Polons −126 ,173 do nStudies in the early 1990s led to the purification, cloning, and sequencing of human PON1 and the molecular characterization of its polymorphisms ,175,176.PON1 modify PON1 properties, and some SNPs can functionally affect the concentration and/or the activity of PON1 [Human PON1 has nearly 200 SNPs ,178. Of  of PON1 ,180,181. of PON1 ,183,184.PON1 coding region imply the catalytic activity of PON1 in the hydrolysis of specific substrates [It has been shown that the SNPs of the bstrates ,179,185.bstrates ,194,195.PON1 55M isoform is associated with less enzymatic activity than the PON1 55L isoform [PON1 55M [The  isoform . HoweverPON1 55M  and/or tPON1 55M .PON1 Q192R polymorphism due to the two isoforms, which differ considerably in their affinity for and catalytic activity with many substrates [PON1 Q192R polymorphism affects the catalytic efficiency of PON1 toward some of its substrates [PON1 192R isoform hydrolyzes paraoxon (an active metabolite of the parathion) up to six times faster than the 192Q isoform, but some organophosphates, sarin, soman, and lactones are hydrolyzed faster by PON1 192Q isoform [More attention has been paid to the bstrates ,198. Thebstrates ,199,200. isoform ,201,202.PON1 gene affect the rate of HDL protection against LDL oxidation [L55M and Q192R polymorphisms and susceptibility to coronary artery disease [It has been demonstrated that polymorphisms of the xidation ,210. How disease ,211. How disease ,212,213.PON1 , diabetes mellitus, metabolic syndrome, aging, and some neurological disorders. Additionally, studies suggested the possible role of PON1 in its development ,233,234.Decreased PON1 activity has been associated with a high risk of CVD . SimilarThe level of PON1 as an antioxidant and antiatherogenic enzyme decreases in diabetes mellitus ,246,247.Additionally, the liver plays a critical role in synthesizing serum PON1, with the gene expression confined only to the liver. Therefore, any injury could influence its status . NonalcoOxidative stress, such as LDL oxidation, is thought to be an important mechanism in many degenerative diseases, including atherosclerosis, diabetes, and AD . FurtherIn epidemiological studies, confounding variables such as environmental and physiological factors should be obtained before analysis. Sex is a biological factor that impacts PON1 expression and activity. In humans, both arylesterase and lactonase activity is increased in women compared to men. The difference was independent of age, HDL-c, and smoking . HoweverIt has been shown that various non-genetic factors can modulate the activity and expression of PON1 through different mechanisms partially understood, involving nuclear receptors, at least, at the transcriptional level. The most prominent factors are diet, drugs, and environmental pollutants ,216,266.PON1 promoter region contains a nonclassical core ARE-like motif between positions −126 and −106, as reported by Gouédard et al. [PON1 expression at the transcriptional level via a JNK/c-Jun signaling pathway [The consumption of fruits, vegetables, and red wine is associated with benefits against oxidative stress diseases, major due to polyphenols. Studies have shown that fruit juices and red wine polyphenols can increase the activity of PON1 both in mice and in humans ,268. Qued et al. . Additiod et al. ,272,273. pathway . The Med pathway .PON1 expression through the cholesterol transcriptional regulator (Sterol response element binding protein 2 (SREBP2)) and Sp1 binding to the PON1 promoter region. The presence of an important SREBP2 (bp –104 to –95) in the proximal PON1 promoter region suggests an important PON1–lipoprotein relationship. Furthermore, SREBP-2 increased the PON1 promoter activity in a dose-dependent manner, indicating an additional lipid-related mechanism of PON1 activation [PON1 synthesis through PPAR activation [PON1 gene are controversial; the increase in PON1 expression by fibrates suggests it does not involve this nuclear receptor. However, in vivo treatment with fenofibrate increased PON1 expression, but the in vitro result of this induction is unclear [PON1 promoter region [PON1 expression by the PPARγ activation [PON1 expression via the PPARγ-PKA-cAMP pathway in Huh7 cells. Additionally, a urokinase-type plasminogen activator (uPA) was observed that decreased PON1 expression by inactivating PPARγ, and this mechanism depended on uPA-mediated mitogen-activated protein kinase activity [PON1 expression in a time-dependent GR activation; the interactions of the response element–GR ligand complex in the PON1 promoter region were confirmed by chromatin immunoprecipitation between −95 and −608 base pairs of the promoter region [It has been demonstrated that statins, glucose, and quercetin induce tivation . Additiotivation . However unclear ,153, sinr region . Similartivation ,278,279.activity . Howeverr region .PON1 promoter region [PON1 expression is limited, and the mechanism involved in PON1 regulation is still unclear. The protective effect of PON1 on dichlorvos-induced liver oxidative stress injury in mice has been studied. The upregulation in PON1 reduced malondialdehyde content, enhanced superoxide dismutase (SOD) and catalase (CAT) activities, increased GSH content, and may have also upregulated Nrf2 mRNA expression [PON1 expression, since Wagner et al. [PON1 promoter region. Moreover, hepatocytes incubated with roasted curcumin or 4-vinyl guaiacol showed enhanced transactivation of Nrf2 and the induction of PON1 [PON1 transcriptional regulation through Nrf2 are not entirely understood. Moreover, several studies have reported the decrease in PON1 activity due to oxidative stress-mediated inflammatory reactions in organophosphate (OP) exposure [The study of the effect of environmental pollutants on PON1 has been focused mainly on pesticides. It has been reported that putative binding sites for PXR exist in the r region , and stur region . Howeverpression . This sur et al.  identifi of PON1 . Howeverexposure ,287,288.exposure . Low actexposure .With the current breakthrough in omics platforms, environmental exposure in utero and during the early stages of development can cause permanent changes through epigenetic programming, leading to chronic degenerative diseases as severe as cancer.Therefore, we consider the definition of current epigenetics, involving molecular modifications in DNA that can regulate gene activity independent of the DNA sequence, whose changes are, in turn, mitotically stable . PreviouThe relationship between the social environment and health has been established in epidemiology. The social  and physical  dimensions are involved in this context concerning the environment; factors that interact together regarding an organism, determining its development and survival ,294. FurPON1 has been cataloged as a model for integrating genetic and epigenetic data due to its multifunctional enzyme involved in oxidative defense, with its genetic variants and lower levels associated with adverse health effects, such as alterations in neurodevelopment and at the cardiometabolic level .PON1 gene is highly polymorphic and more than 400 SNPs identified in the coding region, introns, and regulatory region have been reported. However, many of these SNPs remain unidentified, but the SNPs studied and identified have presented a direct relationship with the activities of PON1 [Age and different exogenous and endogenous factors, and genetic context, are potential factors associated with PON1 variability and susceptibility through epigenetic regulation and the study of different mechanisms in this area; the relationship with the presence of different diseases can be explained . The PON of PON1 .PON1 gene is a member of a group of genes that include PON1, PON2, and PON3 and remain adjacent to each other located on chromosome 7. Specifically, the PON1 promoter region has a CpG island comprising 19 CpG sites, and a second island mapped near exon 7 contains eight CpG sites. Overall, 287 CpG sites were reported in PON1 [The  in PON1 .PON1 gene have focused on the varying DNA methylation levels. Changes in methylation profiles, specifically at these sites, may lead to potential effects at the gene expression level due to the specific binding of transcription factors at these CpG sites; however, epigenetically, data on the methylation of these sites are scarce [PON1 have been evidenced in the promoter and the coding regions of this gene in populations exposed to several toxic compounds, such as heavy metals and pesticides, and in populations diagnosed with the development of different diseases, including metabolic syndrome, ischemic stroke, and coronary artery disease, among others [So far, most of the studies on epigenetic modifications in the e scarce . Hypo- ag others ,298,299.PON1 and the development of cardiovascular disease. Interestingly, it has been identified that most of these DMPs are located in the promoter region, CpG islands, and in the transcription factor-binding sites, suggesting a possible direct link with gene expression and, in turn, some genes that relate to these DMPs have been associated with several neuroendocrine signaling pathways [Regarding epigenome studies, differentially methylated regions (DMRs) and differentially methylated positions (DMPs) have been associated with the presence of specific genotypes of pathways . PON1; however, only one study has evaluated, through an obesity-resistant rat model, the implications of the epigenetic regulation by histone methylation on the expression of this gene. Results at the promoter region show that acetylated histone H4 (H4Ac) and dimethylated histone H3 at lysine residue 4 (H3K4Me2) marks were significantly higher in the high-fat group than the control group [PON1 regulation. However, longitudinal studies are necessary to extend the study of these modifications and their involvement in the possible development of diseases attributed to the loss of PON1 regulation.Additionally, regarding histone modifications, there have been no population studies focused on evaluating the implications of this epigenetic modification on the expression of ol group , suggestHowever, regarding the expression of miRNAs assessed by in silico analyses, several miRNAs have been identified, including miR-616, miR-19a, miR-26a, miR-505, miR-495, miR-153, and miR-185, involved in the negative regulation of PON1 . MoreovePON1 gene. Thus, it has been evidenced in an inverse association between the methylation of CpG sites of the promoter of PON1 with the activity AREase in adults with dietary restriction [PON1, the corresponding data remain scarce in the literature; however, the PON2 and PON3 genes have been studied, focusing on the levels of the methylation profile of these genes  [Regarding non-coding RNAs (ncRNAs), Chen et al.  investigg others ,310,311.atterns) .PON1 has been linked to the development of diseases, such as vascular dementia and cardiovascular diseases, among others. Nevertheless, information about the PON1 gene and DNA methylation in cancer development is still scarce. A study in renal cell carcinoma (RCC) found second differentially methylated imprinted sites related to RCC’s higher methylation levels than normal cells. Of the 15 RCC tumor tissues evaluated, 12 had high methylation of PON1. Additionally, this study demonstrated that the demethylation of PON1 inhibited the migration, invasion, and proliferation of renal cancer cells and arrested more cells in the G0/G1 phase [PON1, which could be used as a targeted biomarker in treating cancers, such as RCC. Moreover, in colorectal cancer (CRC), patients were found with elevated oxidative stress landmarks (DNA damage and advanced oxidation protein products), a decrease in the level of PON1 activity, and an increase in histone deacetylase activity, suggesting the crucial influence of histone deacetylase activity on PON1 in cancer development [Nowadays, the variation of DNA methylation levels of G1 phase . These felopment .PON1 expression is inversely associated with a degree of vascular invasion in hepatocarcinoma cells.Several studies have demonstrated the relationship between oxidative stress and the development of human diseases among these cancers. Recently, many studies have focused on the relationship between PON1 and cancer. In this regard, some studies have reported low expression and activity of PON1  in patients with different types cancer such as cancer blander , gastroiPON1 gene have been implicated in cancer development. The polymorphism of PON1 most studied is PON1 Q192R because the two isoforms differ considerably in their affinity for and catalytic activity with several substrates. This polymorphism has been related to bladder cancer, renal cancer, prostate cancer, and lymphoma (PON1-192R had reduced risk in the development of brain cancer and breast cancer. Concerning PON1 L55M, it is also associated with a higher risk of prostate cancer [PON1 L55M was significantly associated with hematological tumors and breast cancer. Therefore, these authors suggest this polymorphism as a biomarker to susceptibility to identify an individual with a risk of hematological tumors and breast cancer.Additionally, polymorphism genetics of the lymphoma . Howevere cancer  (Table 5e cancer , only 40PON1 polymorphisms regarding cancer association, more studies are required to establish the other PON1 polymorphisms’ contribution to cancer development.Since we did not find information on other PON1 and the relationship between PKC and MAPK/ERK pathways activated by growth factors to regulate cell growth and differentiation, apoptosis, and angiogenesis [Based on the studies, it is evident that PON1 plays a vital role in cancer. However, it is important to account for the cancer type, ethnicity, gene interactions, and several factors contributing to cancer development. Additionally, the extensively signal transduction pathways involved in the modulation of ogenesis .This review involves the role of PON1 and cancer. Undoubtedly, the gene expression of PON1 will remain challenging to study and therefore, more studies at different levels of scientific research are required for the complete elucidation of the role of this enzyme and its specific relationship with cancer. Therefore, targeting PON1, redox-sensitive pathways and transcription factors promise disease prevention, including cancer.


Background: Copper is an indispensable mineral element involved in many physiological metabolic processes. Cuproptosis is associated with a variety of cancer such as hepatocellular carcinoma (HCC). The objective of this study was to examine the relationships between the expression of cuproptosis-related genes (CRGs) and tumor characteristics, including prognosis and microenvironment of HCC.Methods: The differentially expressed genes (DEGs) between high and low CRGs expression groups in HCC samples were identified, and further were analyzed for functional enrichment analysis. Then, CRGs signature of HCC was constructed and analyzed utilizing LASSO and univariate and multivariate Cox regression analysis. Prognostic values of CRGs signature were evaluated by Kaplan-Meier analysis, independent prognostic analysis and nomograph. The expression of prognostic CRGs was verified by Real-time quantitative PCR (RT-qPCR) in HCC cell lines. In addition, the relationships between prognostic CRGs expression and the immune infiltration, tumor microenvironment, antitumor drugs response and m6A modifications were further explored using a series of algorithms in HCC. Finally, ceRNA regulatory network based on prognostic CRGs was constructed.Results: The DEGs between high and low CRG expression groups in HCC were mainly enriched in focal adhesion and extracellular matrix organization. Besides, we constructed a prognostic model that consists of CDKN2A, DLAT, DLST, GLS, and PDHA1 CRGs for predicting the survival likelihood of HCC patients. And the elevated expression of these five prognostic CRGs was substantially in HCC cell lines and associated with poor prognosis. Moreover, immune score and m6A gene expression were higher in the high CRG expression group of HCC patients. Furthermore, prognostic CRGs have higher mutation rates in HCC, and are significantly correlated with immune cell infiltration, tumor mutational burden, microsatellite instability, and anti-tumor drug sensitivity. Then, eight lncRNA-miRNA-mRNA regulatory axes that affected the progression of HCC were predicted.Conclusion: This study demonstrated that the CRGs signature could effectively evaluate prognosis, tumor immune microenvironment, immunotherapy response and predict lncRNA-miRNA-mRNA regulatory axes in HCC. These findings extend our knowledge of cuproptosis in HCC and may inform novel therapeutic strategies for HCC. Hepatocellular carcinoma (HCC) accounts for approximately 90% of liver cancers and is a common cause of death in cancer patients . The WorPrevious studies showed that a variety of therapeutic methods involving programmed cell death such as pyroptosis and ferroptosis play an important role in inhibiting the occurrence and development of tumors . Among tRecently, This study comprehensively investigated the molecular alterations and clinical relevance of CRGs in HCC. A total of 371 HCC patients were selected from The Cancer Genome Atlas (TCGA) database, classified into 2 cuproptosis-related subtypes by consensus clustering based on 19 CRGs, the signaling pathways of differential gene enrichment in the two subtypes were analyzed, and the differences in immune cell infiltration, important immune checkpoints, and m6A methylation-related genes between the two subtypes were measured. The CRGs associated with the prognosis of HCC patients were also analyzed by logrank test and univariate regression analysis. Gene and protein expression of prognostic CRGs in HCC cells or tissues were also assessed, as well as their association with anti-tumor drug sensitivity, m6A methylation-related genes, and tumor microenvironment (TME). Prognostic models were developed to predict the overall survival (OS) and progression-free survival (PFS) in patients with HCC. A competing endogenous RNA (ceRNA) regulatory network was constructed to screen the lnRNA-miRNA-mRNA network that affected the prognosis of HCC patients. Our analysis highlighted the importance of CRGs in HCC development and laid the foundation for the therapeutic application of cuproptosis regulators in HCC.https://portal.gdc.cancer. Gov/)  . A totalNineteen CRGs were first extracted from the TCGA expression matrix. According to the consistent clustering of these 19 genes, the R software package Consensus-ClusterPlus (v1.54.0) was used for the analysis of consistency, and the maximum number of clusters was 6 . The TCGp values were analyzed in the TCGA database to correct for false positive results. “Adjusted p < 0.05 and log2 (fold change) > 1 or log2 (fold change) < −1″ was defined as the criteria for screening the differential expression of mRNA. GeneMANIA (http://www.genemania.org), which is a software that elucidates the relationship between genes and datasets by building gene interaction networks (DEGs between C1 and C2 subgroups were obtained using Limma package (version 3.40.2) in R software . Adjustenetworks , was usehttp://software.broadinstitute.org/gsea/index.jsp)  < 0.25. Statistical analysis and graphing were performed using the R package clusterProfiler (3.18.0).The GO function and the enrichment of KEGG pathways were analyzed by the “clusterProfiler” package in R software . Potentidex.jsp) . AccordiThe R software package immunedeconv was used for immune score assessment to compare the degree of immune cell infiltration in C1 (N = 252) and C2 (N = 119) subgroups by Wilcoxon test, by integrating six state-of-the-art algorithms, including TIMER , xCell , MCP-coup values, and hazard ratios (HR) with 95% confidence intervals (CIs) were obtained by logrank test and univariate Cox regression. Five CRGs  with higher hazard ratios were screened from the Cox regression analysis plot. The relationship between the prognostic CRGs and the OS rate in HCC patients was also analyzed, and the AUC (Area Under Curve) under the ROC curve was calculated.The expression of CRGs in 371 HCC tissues and 50 paired adjacent tissues was analyzed using the TCGA database. In addition, univariate Cox regression analysis was used to investigate the effect of CRGs on the prognosis of HCC. Kaplan-Meier curves, 2.The following HCC cell lines and hepatocyte were used in this study: Hep3B, Huh7, HepG2, and L02 . All cell lines were maintained in Dulbecco’s modified Eagle’s medium  supplemented with 10% fetal bovine serum , 100 U/ml penicillin, and 100 U/ml streptomycin  and incubated at 37°C in a humidified atmosphere with 5% CO−△△CT) was employed to determine the relative mRNA expression, with the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene as the reference. First, TRIzol reagent  was used to extract total RNA from the samples. Then, the total RNA concentrations were measured using a NanoDrop 2000c . Next, cDNA was synthesized by reverse transcription using a miScript II RT kit . Finally, a miScript SYBR Green PCR kit  was used to detect the expression of target genes on a Lightcycler 480 Real-Time PCR System . The 20-μl reaction mixture included 10 μl 2X QuantiTect SYBR Green PCR Master Mix, 0.8 μl 10X miScript universal Primer, 2 μl cDNA template, 0.8 μl specific miRNA primer, and 7.4 μl RNase-free water. Additionally, the thermocycling steps were as follows: 95°C for 15 min, followed by 40 cycles of 94°C for 15 s, 55°C for 30 s, and 72°C for 30 s. The relative standard curve method  was displayed by univariate and multivariate cox regression analysis and using forest plots by the “forestplot” package. The “rms” package was used to build a Nomogram model for predicting 1, 3, and 5-year OS and PFS based on the results of multivariate cox proportional hazards analysis.A prognostic model was constructed using LASSO-Cox regression analysis based on the above CRGs associated with the prognosis of HCC patients. According to the results of the multivariate Cox regression analysis, the prognostic CRG risk score was calculated as follows: Risk score = ∑i Cofficient(mRNAi) × Expression (mRNAi). Then, TCGA-LIHC patients were divided into low-risk and high-risk subgroups according to the average risk score, Kaplan-Meier analysis was used to compare the OS rates of the two subgroups, and timeROC analysis was performed to predict the accuracy of the model. Each variable (including the http://gepia2.cancer-pku.cn/#index). The UALCAN database  is an interactive portal containing TCGA RNA transcriptome data and clinical data from 31 cancer types. TISIDB (http://cis.hku.hk/TISIDB) is a portal for assessing tumors and the immune system that integrates multiple heterogeneous data types including genomics, transcriptomics, and clinical data from TCGA of 30 cancer types  database to assess the differences in CRG expression between HCC and adjacent tissues  provides visualization tools for analyzing cancer genetic data. The genomic maps of CRGs were analyzed using cBioPortal based on the TCGA database to understand the mutation frequency in HCC  database   . Next, thttps://www.cancerrxgene.org/)  . The R pThe correlation between 19 CRGs and m6A-related genes expression and prognostic in 371 HCC samples was analyzed by R (4.0.3) packages “ggplot2.” The difference in m6A-related gene expression between C1 group (N = 252) and C2 group (N = 119) was investigated. The m6A-related genes analyzed included METTL3, YTHDC1, YTHDC2, METTL14, RBM15, RBM15B, IGF2BP1, IGF2BP2, IGF2BP3, VIRMA, WTAP, YTHDF1, YTHDF2, YTHDF3, ZC3H13, HNRNPA2B1, HNRNPC, RBMX, FTO, and ALKBH5.http://tisch.comp-genomics.org/) (The effect of prognostic CRGs on the expression of single cell subsets in the TME was investigated using TISCH (cs.org/) . TISCH ihttp://starbase.sysu.edu.cn/) database (http://www.rnainter.org/) (https://cm.jefferson.edu/rna22/interactive) database (http://www.mirnet.ca/) database (http://www.cytoscape.org/) software   and RNA2database , and thedatabase . Subsequsoftware . In addiThe flowchart of the study is illustrated in DLD, GLS, NFE2L2, DLST, LIAS, ATP7B, PDHA1, SLC31A1, FDX1, DBT, GCSH, LIPT1, CDKN2A, DLAT, NLRP3, ATP7A, PDHB, LIPT2, and MTF1) were related to several processes, including oxidoreductase complex, tricarboxylic acid cycle enzyme complex, cellular amino acid catabolic process, dihydrolipoyl dehydrogenase complex, acyl-CoA metabolic process, acetyl-CoA biosynthetic process and thioester metabolic process , HAVCR (p < 0.01), PDCD1LG2 (p < 0.01), TIGIT (p < 0.05) and SIGLEC15 (p < 0.05) were highly expressed in the C1 subgroup than in the C2 subgroup , Macrophage M2, B cell ; Macroph NK cell ; NK cell NK cell ; Macrophutrophil ; MacrophCD4+ Th1 ; T cell tic cell  respectisubgroup .ATP7A, SLC31A1, LIAS, LIPT1, LIPT2, DLD, DLAT, PDHA1, PDHB, MTF1, GLS, CDKN2A, and DLST was upregulated, while NLRP3 and DBT were downregulated in cancer tissues than in the normal tissues (p < 0.001) in HCC cell lines  compared to their expression in the hepatocyte cell line (L02) , DLAT , DLST , GLS  and PDHA1  were highly expressed.The expression of 19 CRGs in HCC and normal tissues was investigated using the TCGA dataset; among them, 15 CRGs were up- or downregulated. The expression of  tissues . Most of tissues . Univari tissues . The mRNne (L02) . FinallyGLS, DLAT, and CDKN2A   . The are0.00106) . The resCDKN2A expression and pT stage were independent factors affecting the prognosis of HCC patients , while the correlation between DLST and the pathological stage was not significant  2 database. The results showed that the expression of nificant . The anaficantly .The protein expression of CRGs in 371 HCC and adjacent tissues was measured using the Human Protein Atlas to determine the difference in protein expression of the prognostic CRGs in HCC tissues and normal liver. The results of the immunohistochemical staining showed that the prognostic CRGs were moderately or highly expressed in HCC tissues but were low in paracancerous tissues .CDKN2A, DLAT, DLST, GLS, and PDHA1 were altered in 80%, 25%, 48%, 37%, and 32% of the 347 samples of the 372 sequenced patients, as shown in p = 0.0377, p = 0.018) of HCC patients. However, these two groups have little correlation with OS  online tool for HCC. CDKN2A, DLAT, DLST, GLS, and PDHA1) and immune infiltration in HCC using the TIMER database and TCGA database. TIMER data showed that CDKN2A, DLAT, GLS and PDHA1 were strongly and positively correlated with tumor purity, B cells, CD4+ T cells, CD8+ T cells, neutrophils, macrophages, and dendritic cells, while DLST was only positively correlated with tumor purity, CD4+ T cells, neutrophils, and dendritic cells , PD-L1  and CTLA-4 , and GLS was also positively correlated with CD274 , PD-L1  and CTLA-4 , while DLAT , DLST  and PDHA1  were significantly and positively correlated only with CD274  (CDKN2A (p = 1.64 × 10−04) was positively correlated with TMB, and GLS (p = 7.74 × 10−05) and negatively correlated with TMB  and TMB can be used as indicators to predict the response of various tumors to immunotherapy . The abo= 0.044) . Furtherwith TMB . Finallycleotide .IGF2BP1, other m6A-related genes were significantly different between the two groups  were found between the C1 and C2 subgroups. The results showed that except http://tisch.comp-genomics.org/) to examine the relationship between prognostic CRGs and the infiltration of immune cells, stromal cells, and malignant cells. The HCC single-cell GSE dataset was explored (LIHC_GSE125449_aPDL1aCTLA4). T cells, B cells, plasma cells, macrophages, endothelial cells, fibroblasts, malignant cells, and hepatic progenitors were annotated by single-cell RNA sequencing analysis , myofibroblasts, immune cells and other factors. The prognostic CRGs were assessed at a single-cell level expression by Tumor Immunity Single Cell Center , 3 miRNAs  and 6 lncRNAs . Eight lncRNA-miRNA-mRNA networks were also constructed, including lncRNA CYTOR/miR-125-5p/CDKN2A, lncRNA CYTOR/miR-125-5p/GLS, lncRNA DANC R/miR-125-5p/GLS, lncRNA DANCR/miR-125-5p/PDHA1, lncRNA DLEU2/miR-23c/GLS, lncRNA GSEC/miR-101-3p/GLS, lncRNA PTPRG-AS1/miR-101-3p/GLS and lncRNA TRAF3IP2-AS1/miR-23c/GLS regulatory axes. . The results of prognostic analysis showed that HCC patients with higher expression of CDKN2A, DLAT, DLST, GLS, and PDHA1 had shorter OS, indicating that prognostic CRGs promoted the progression of HCC. CDKN2A is highly expressed in various cancer tissues such as liver cancer and kidney cancer and plays an important prognostic role in many cancers . DLST isGLS, DLAT, and CDKN2A increased the risk score, and the OS rate of patients in the high-risk group was significantly shorter than that of the patients in the low-risk group. In addition, CDKN2A was found as an independent risk factor, and a valid nomogram was constructed to predict the 1-, 3-, and 5-year survival rates of HCC patients, suggesting that CDKN2A played an important role in the occurrence and prognosis of HCC.Univariate Cox regression and LASSO Cox regression analysis were subsequently used to analyze the five signature genes. The prognostic value of these five genes was then tested, and the results showed that HCC patients with high expression of these five CRGs had decreased survival rates. Moreover, the higher expression of Most malignancies are caused by somatic mutations within the cancer genome, and mutational signatures correlate with mRNA and protein expression in multiple cancer types . Functio+/low CD20+ immune cell infiltration ratio is an important prognostic factor for lung squamous cell carcinoma (+ T cells expressing the chemokine receptor 7 [T(ccr7)], demonstrating that patients with colorectal cancer with high T (ccr7) and T (reg) invasion have a better prognosis . Corognosis . In the At present, immunotherapy targeting immune checkpoints, especially programmed cell death protein 1/programmed cell death ligand 1 (PD-1/PD-L1) and CTLA-4 blockers have become feasible in the treatment of many malignant tumors . PD-1 isCDKN2A increased the TMB score in HCC and PDHA1 increased the MSI score. Furthermore, prognostic CRGs were positively or negatively associated with multiple chemotherapeutic agents. These results provide new potential therapeutic targets in the treatment of HCC.The research on tumor immune checkpoint inhibitors in tumor immunotherapy is the most developed. TMB and MSI are considered potential predictive biomarkers involved in response to ICIs . High TMvia METTL3-mediated MYC mRNA m6A modification (N6-methyladenosine (m6A) is the most prevalent internal mRNA modification in mammalian cells. RNA methylation is a pervasive post-transcriptional modification that plays a key role in regulating various biological processes, and its dysregulation is closely related to the occurrence of human malignancies  through fication . CurrentStromal components, including CAFs and TAMs, play important roles in cancer initiation and progression . Our resEight lncRNA-miRNA-mRNA networks were also constructed, lncRNA DANCR reduces the expression of miR-125b-5p and regulates the proliferation and apoptosis of hepatoma cells . LncRNA + T cells, promoted the expression of immune checkpoints such as CD274 and HAVCR2, and promoted the expression of various m6A-related genes in HCC. CRGs also participated in multiple cancer-related signaling pathways in HCC. A total of 19 CRGs that were highly expressed in HCC were analyzed, and five CRGs  that were associated with the prognosis of HCC patients were identified. A prognostic model of CRGs was established, which predicted more accurately OS and PFS in HCC patients. Mutations in prognostic CRGs led to poorer prognosis in patients with HCC. Prognostic CRGs were positively correlated with B cells, T cells, macrophages, and other immune cells, and were positively correlated with various chemotherapeutic drugs. They affected the TME in HCC by affecting CAFs and TAMs. Finally, eight lncRNA-miRNA-mRNA regulatory axes that affected the progression of HCC were predicted. Hence, our study laid the foundation for an in-depth understanding of the role of CRGs in HCC.CRGs affected the infiltration of immune cells such as macrophages and CD8


PPO encoding genes, PPO1 and PPO2. To date, no PPO gene or mutant has been characterized in monocotyledonous plants. In this study, we isolated a spotted and rolled leaf (sprl1) mutant in rice (Oryza sativa). The spotted leaf phenotype was sensitive to high light intensity and low temperature, but the rolled leaf phenotype was insensitive. We confirmed that the sprl1 phenotypes were caused by a single nucleotide substitution in the OsPPO1 (LOC_Os01g18320) gene. This gene is constitutively expressed, and its encoded product is localized to the chloroplast. The sprl1 mutant accumulated excess Proto(gen) IX and reactive oxygen species (ROS), resulting in necrotic lesions. The expressions of 26 genes associated with tetrapyrrole biosynthesis, photosynthesis, ROS accumulation, and rolled leaf were significantly altered in sprl1, demonstrating that these expression changes were coincident with the mutant phenotypes. Importantly, OsPPO1-overexpression transgenic plants were resistant to the herbicides oxyfluorfen and acifluorfen under field conditions, while having no distinct influence on plant growth and grain yield. These finding indicate that the OsPPO1 gene has the potential to engineer herbicide resistance in rice.Protoporphyrinogen IX (Protogen IX) oxidase (PPO) catalyzes the oxidation of Protogen IX to Proto IX. PPO is also the target site for diphenyl ether-type herbicides. In plants, there are two  Higher plants produce four classes of tetrapyrroles, including siroheme, heme, phytochromobilin, and chlorophyll (Chl). Tetrapyrroles are synthesized via a multibranched pathway. All four classes of tetrapyrroles share the six biosynthesis steps from glutamate to uroporphyrinogen III. The three subsequent steps from uroporphyrinogen III to protoporphyrin IX (Proto IX) are common to Chl, heme, and phytochromobilin synthesis. Finally, Fen of Chl . So far,bidopsis ,2. Howevbidopsis ,13,14,15Arabidopsis, tobacco (Nicotiana tabacum), spinach (Spinacia oleracea), and other plants, which share only 24% and 27% identity in Arabidopsis and tobacco, respectively [hemG gene encoding PPO was first identified from Escherichia coli (E. coli) [PPO genes have been characterized from human, yeast, and plants [Arabidopsis, tobacco, and spinach PPO genes have been isolated by the functional complementation of the E. coli hemG mutant [Arabidopsis, transgenic plants expressing the antisense PPO1 gene showed the necrotic lesions phenotype, homozygous ppo1 null mutants from T-DNA insertion displayed etiolated cotyledons and a seedling lethal, and heterozygous ppo1-1/+ plants showed reproductive growth defects [PPO1 antisense plants showed a reduced growth rate and necrotic leaf damage and generated more necrotic leaf lesions under low-light than under high-light exposure [Protoporphyrinogen IX (Protogen IX) oxidase (PPO) catalyzes the oxidation of Protogen IX to form Proto IX, which is the last common step in Chl and heme biosynthesis. There are two genes encoding PPO proteins (PPO1 and PPO2) in ectively ,17,18,19ectively ,17. The E. coli) . Up to nd plants ,21,22. IG mutant ,17,18. I defects ,24,25. Iexposure .Bacillus subtilis and Arabidopsis PPO genes in tobacco generated resistance to the DPE herbicides oxyfluorfen and acifluorfen, respectively [Bacillus subtilis, Arabidopsis, human, and Myxococcus xanthusPPO genes were resistant to oxyfluorfen under controlled growth conditions [PPO is a molecular target site for photodynamically active herbicides of the diphenyl ether (DPE) type, which inhibits PPO activity by competing with its substrate Protogen IX. When DPE interacts with PPO, excess Protogen IX diffuses from chloroplasts into the cytoplasm and is oxidized to photosensitizing Proto IX by an unspecific plasma membrane-bound peroxidase. Proto(gen) IX absorbs excessive light energy, which finally results in the formation of highly reactive oxygen species (ROS). Then, ROS provoke the rapid degradation of membranes, proteins, and DNA and eventually lead to cell death ,28. The ectively . Transgenditions ,33,34,35PPO gene or mutant has been reported in monocotyledonous plants, and the function of PPO in rice remains to be explored. In the present study, we isolated a spotted and rolled leaf (sprl1) mutant from the ethyl methanesulfonate (EMS)-treated indica rice restorer line Lehui188 (188R). The sprl1 mutant exhibited necrotic lesions on leaves throughout development and extremely inwardly rolled leaves before the three-leaf stage. We cloned the mutated gene and confirmed that the mutant phenotype of sprl1 was caused by a single nucleotide substitution in LOC_Os01g18320 gene encoding OsPPO1. The sprl1 mutant accumulated excess Proto(gen) IX and ROS. Unlike the PPO1 antisense tobacco plants, the sprl1 leaves displayed more necrotic lesions under high light intensity or low temperature. Importantly, the overexpression of the OsPPO1 gene generated resistance to the DPE herbicides oxyfluorfen and acifluorfen under field conditions, while having no detrimental effect on plant growth or grain yield, indicating the application potential of OsPPO1 in rice.Nonetheless, no sprl1 mutant displayed reddish-brown lesions on the leaves throughout the whole growth period , 3,3′-diaminobenzidin (DAB), and trypan blue (TB) staining. In NBT and DAB staining, the leaves with lesions were dyed deep blue and brownish in the mutant, respectively, but no staining was observed in the wild type leaves , superoxide dismutase (SOD), and peroxidase (POD). The results showed that the activities of SOD and POD in the sprl1 leaves were remarkably elevated by 89.8% and 129.5%, respectively, compared to those in the wild type  is a product of membrane lipid peroxidation and an indicator of free radical production and consequent tissue damage . So, we ild type G,H, whilild type F.sprl1 mutant with its wild type parent 188R. The resulting F1 plants all displayed a normal leaf phenotype. The F2 populations exhibited the segregation of the wild type plants and the mutant plants at the seedling stage, with a ratio of 3:1 , suggesting that the spotted and rolled leaf phenotype of sprl1 is controlled by a single recessive nuclear gene.For genetic analysis of the mutant phenotype, we crossed the sprl1, we carried out whole-genome resequencing and a MutMap analysis. A bulked DNA pool from 30 mutant plants showing spotted and rolled leaves in the (sprl1 × 188R) F2 population was sequenced and analyzed. Consequently, a linkage region was identified on the short arm of chromosome 1 (LOC_Os01g18320 (gene1) and LOC_Os01g22954 (gene2), whose SNP-index values were 1.0 and 0.92, respectively. Meanwhile, to further identify the sprl1 mutant gene, we further analyzed 353 homozygous recessive individuals from the (sprl1 × japonica cultivar Zhonghua 11) F2 mapping population, using five insertion/deletion (InDel) markers within the vicinity of the two gene loci  in tetrapyrroles biosynthesis. We amplified and sequenced LOC_Os01g18320 in the sprl1 mutant and its wild type, respectively. The result confirmed that LOC_Os01g18320 carries a single nucleotide C-to-T substitution at position 516 in the sprl1 mutant, consistent with the MutMap analysis. cDNA sequencing analysis showed that the C-to-T substitution occurred at position 427 in the cDNA of this gene . The BlastP results showed that there is one homologous gene (LOC_Os04g41260) of OsPPO1 in rice, which is predicted to encode OsPPO2. OsPPO2 shares 27% identity with OsPPO1. While the OsPPO2 protein is predicted to be targeted to the mitochondria, the OsPPO1 protein contains a chloroplast-targeting sequence of 36 amino acid residues at its N terminus , barley (Hordeum vulgare), Arabidopsis, potato (Solanum tuberosum), and tobacco, with identities of 89, 88, 71, 70 and 69% respectively, but it only has low identities to the PPOs of other organisms such as Bacillus subtilis, Homo sapiens, and Myxococcus xanthus . A phylogenetic analysis showed that OsPPO1 is more closely related to the homologs from monocotyledon maize and barley than to those from other species  IX, confirming that the mutant indeed has a defective PPO1 protein.Because Protogen IX is the substrate for PPO1 in tetrapyrroles biosynthesis, there should be an excessive accumulation of Protogen IX and photo-oxidized Proto IX when the plants have a defect in the PPO1 protein . For a qd method . The resl g−1 FW . Therefosprl1 was caused by the mutation of the OsPPO1 gene, we performed a complementation analysis. The construct pCAMBIA1300-OsPPO1, containing a 5528 bp OsPPO1 genomic DNA fragment consisting of a 1949 bp upstream sequence, the entire coding region, and a 333 bp downstream sequence, was introduced into the sprl1 mutant by Agrobacterium-mediated transformation. As a result, 18 independent transgenic lines were successfully obtained. Among them, 11 lines were identified as positive transgenic lines by the PCR examination and sequencing analysis, which displayed normal phenotypes compared to the wild type, including the leaf blade, plant height, and other major agronomic traits (OsPPO1 (LOC_Os01g18320) is responsible for the spotted and rolled leaf phenotype of sprl1.To further confirm that the mutant phenotype of c traits . Furtherc traits D–F. OverOsPPO1-GFP and p35S:GFP (as control) in rice protoplasts were observed using laser scanning confocal microscopy. As shown in OsPPO1 was predicted to contain a chloroplast transit peptide with 36 amino acid residues at its N-terminus by TargetP and ChloroP . To detePPO1 antisense RNA displayed more necrotic lesions on their leaves under low- compared to under high-light conditions [sprl1 mutant has a similar phenotype as the antisense tobacco plants, we treated the sprl1 mutant and its wild type using various light intensities, including high light , low light , and dim light . As a result, the sprl1 mutant displayed more necrotic lesions under HL conditions than under LL conditions, and no obvious necrotic lesion was observed under DL conditions  under LL conditions, respectively. Consequently, the sprl1 mutant showed severe necrotic lesions under 22 °C conditions, but only a few necrotic lesions were observed under 30 °C conditions  IX and ROS, we examined the transcript levels of the 23 related genes at the seedling stage. Among them, 12 genes are involved in tetrapyrrole biosynthesis, including HEMA (glutamyl-tRNA reductase), GSAM, HEMB (5-aminolevulinate dehydratase), CHLD, CHLH, CHLI, PORA, DVR, YGL, CAO1, FC1, and FC2 (ferrochelatase) [psaA and psbA (two reaction center polypeptides) and rbcL and rbcS  [catA and catB , APX1 and APX2 (ascorbate peroxidase), AOX1a and AOX1b , and POD1 (peroxidase) [OsCHLI and YGL, the expression levels of the genes associated with tetrapyrrole synthesis and photosynthesis were significantly decreased, but the expressions of the genes associated with ROS accumulation were all dramatically increased in the sprl1 mutant  IX and ROS, and necrotic leaves.Since elatase) ,12,39,40subunit) . Seven goxidase) ,43,44,451 mutant B. These sprl1 displayed a leaf rolling phenotype before the three-leaf stage, we also assayed the transcript levels of leaf rolling-related genes in the second leaf blade at the two-leaf stage and the fifth leaf blade at the five-leaf stage, respectively, including ROC5 (Rice Outermost Cell-specific gene5), ZHD1 (zinc finger homeodomain class homeobox transcription factors), REL1 (Rolled and Erect Leaf 1), REL2 (Rolled and Erect Leaf 2), and SRL1 (SEMI-ROLLED LEAF1) [ZHD1, REL1, and REL2 returned to the wild type levels, even as ROC5 and SRL1 were significantly up-regulated in the fifth leaf blade (sprl1.Because D LEAF1) ,48,49,50af blade C. This rOsPPO1, we phenotyped rice plants overexpressing the OsPPO1 gene. The overexpression vector pCAMBIA2300-OsPPO1, containing the full-length cDNA sequence of the OsPPO1 gene under the control of the CaMV 35S promoter, was constructed and then transformed into japonica cultivar Zhonghua 11 (ZH11). Consequently, 16 positive transgenic lines were obtained, in which 7 lines showed an extremely significant increase in the expression level of OsPPO1 compared with the control ZH11 (OsPPO1 overexpressing (OE) lines were indistinguishable from the ZH11 control plants, both with a normal leaf development and growth rate  mutant in rice and identified the causal OsPPO1 (LOC_Os01g18320) gene by a MutMap cloning approach and functional complementation. In the sprl1 mutant, a C-to-T substitution occurred in OsPPO1, resulting in an amino acid change in its encoded protein. The HPLC analysis showed that excess Proto(gen) IX was accumulated in sprl1. Moreover, the sprl1 mutant phenotype can be rescued by complementation with the wild type OsPPO1 gene. Therefore, we successfully identified the PPO1 gene in rice and confirmed that a single nucleotide substitution in this gene caused the spotted and rolled leaf phenotype of sprl1.To date, d plants ,21,22. I spinach ,17,18. HArabidopsis plants expressing the antisense AtPPO1 gene showed a lesion-mimic phenotype, and tobacco plants expressing the antisense NtPPO1 mRNA displayed necrotic leaf damage and a reduced growth rate [sprl1 mutant also displayed a lesion mimic phenotype throughout the growth period (OsPPO1 in sprl1 caused the excess accumulation of photoreactive Proto(gen) IX, resulting in the over-accumulation of ROS, an increased content of MDA, and, eventually, cell death. However, unlike the above mutants of Arabidopsis and tobacco, the sprl1 mutant displayed extremely inwardly rolled leaves at the seedling stage, which resulted from the decrease in the number and size of the bulliform cells in its leaf blades  conditions, but they grew more slowly and displayed more necrotic lesions under low light (LL) conditions [sprl1 mutant had more necrotic lesions under HL conditions but had only a few necrotic lesions under LL conditions and had no necrotic lesion under dim light (DL) conditions. Among these plants, HL-grown sprl1 plants had the highest content of Proto IX and the lowest contents of photosynthetic pigments  IX under LL, DL, and high temperature conditions (sprl1 was not converted into photosensitive Proto(gen) IX, so insufficient levels of photosensitive Proto(gen) IX could induce cellular damage under these conditions. Collectively, the photodynamic damage induced by Proto(gen) IX positively correlates with light intensity but negatively correlates with temperature in rice.Accumulating light-absorbing Proto(gen) IX evokes the generation of ROS and peroxidized lipids. However, the transgenic tobacco plants expressing the antisense nditions . In contpigments . Similarnditions . These dnditions C, indicaPPO genes. For example, transgenic rice expressing Bacillus subtilis PPO was found to be resistant to oxyfluorfen with respect to ion leakage, Chl loss, and lipid peroxidation. While most of these transgenic lines appeared to be normal, morphologically, their seed production varied [Arabidopsis PPO1 or human PPO could germinate in the presence of 20 µM oxyfluorfen, whereas the wild type rice seeds could not [PPO gene showed growth retardation and the light-dependent formation of necrotic lesions [OsPPO1 displayed no phenotypic differences when compared to wild type ZH11 plants, including in terms of photosynthetic pigment contents, plant growth, grain yield, etc.  mutant was isolated from the indica rice restorer line Lehui188 (188R) by ethyl methanesulfonate (EMS) mutagenesis. The rice materials were grown during the natural rice growing season in the experimental farms of Sichuan Agricultural University in Wenjiang District, Chengdu city, China [−2 s−1), low light , and dim light  at 28 °C/12 h of dark at 26 °C, respectively. For temperature treatments, the rice seedlings were grown in the growth chamber at constant temperatures of 30 °C and 22 °C, respectively, under 12 h of low light /12 h of dark.The y, China . For ligsprl1 mutant and wild type at the two-leaf stage were harvested and fixed with a 70% formalin acetic acid–alcohol solution and then dehydrated in a gradient ethanol series. Subsequently, the samples were embedded in paraffin and cut into 8 μm-thick sections. After that, the sections were deparaffinized with xylene and washed in serial dilutions of ethanol. Finally, they were stained with safranin, destained with ethanol, stained with fast green, and observed with a microscope [sprl1 mutant and its wild type at the seedling stage. Then, the leaves were treated as follows: fixed in 3% glutaraldehyde, further fixed in 1% osmium tetroxide, dehydrated, embedded, sectioned, and stained. Finally, sections were observed under a JEM-1400FLASH transmission electron microscope .For paraffin sectioning, the second leaf blades of the croscope . For thesprl1 mutant and its wild type were harvested from plants grown under normal conditions at the five-leaf stage. For the 3,3′-diaminobenzidin (DAB) or nitro blue tetrazolium (NBT) staining analysis, the samples were incubated in DAB or NBT staining solutions at 28 °C in the dark for 12 h, cleared in 95% ethanol, and then photographed [The fresh leaf blades of the ographed . For theographed . The consprl1 mutant and its wild type at the seedling stage and the heading stage, respectively. The pigments were extracted from 0.2 g of fresh leaves with 80% acetone in the dark at 4 °C for 48 h. The contents of chlorophyll (Chl) and carotenoids (Caro) were measured at wavelengths of 470 nm, 646 nm, and 663 nm using the BIOMATE 3S UV-Visible Spectrophotometer  and were calculated according to the method of Lichtenthaler and Wellburn [For the pigment analysis, the leaves were collected from the Wellburn :Chl a (mv/v/v), and the homogenates were centrifuged at 7197 g  at 1 °C for 20 min. To oxidize the Protogen IX into Proto IX, 5 μL of 1 M acetic acid and 5μL of 2-butanone peroxide was added to 200 μL supernatant [For the Proto IX analysis, 0.1 g fresh leaves at the seedling stage were homogenized in 1 mL methanol/acetone/0.1 M NaOH . The resulting plasmid pCAMBIA1300-OsPPO1, which contained a 1949 bp upstream sequence, an entire coding region, and a 333 bp downstream sequence of OsPPO1, was introduced into the sprl1 mutant by Agrobacterium-mediated genetic transformation. The transgenic plants were detected using the primers OsPPO1-Tra F/R, locating on the OsPPO1 gene and the pCAMBIA1300 vector, respectively . The transgenic plants were detected using the primers OSPPO1-OE F2/R2, locating on the OsPPO1 gene and the pCAMBIA2300-35S vector, respectively  were transformed into the 188R protoplasts and incubated overnight in the dark [The full-length cDNA sequence of -GFP F/R , digestethe dark . The gre3 G, analytikjena, Jena, Germany). The Actin 1 gene was chosen as a reference gene. Three biological replicates were carried out in each experiment, and Student’s t test was used for the statistical analysis. All of the primers used in the qRT-PCR analysis are listed in The total rice RNA was extracted from the roots, stems, leaves, sheaths, and young panicles with an RNA isolater kit (Vazyme), and the first-strand cDNA was reverse transcribed using a reverse transcription kit (Vazyme). The quantitative real time RT-PCR (qRT-PCR) analysis was conducted with a real-time PCR system  or 600 μM acifluorfen in a paddy field at sunny-day-dusk. For the germination test, the seeds of OE2, OE15, and the control cultivar ZH11 were surface-sterilized, sown on a half-strength Murashige and Skoog (MS) solid medium containing different concentrations of technical-grade oxyfluorfen  or acifluorfen , and then grown in the growth chamber at 25 °C for 10 days under continuous light after dark incubation overnight . For thet-test, with a p-value less than 0.05 or 0.01 considered significant.Excel in Office 2016 was used to perform the statistical analysis. All of the experiments were repeated independently for three replicates, and the data were subjected to statistical analysis using Student’s


Polyscias balfouriana cv. Marginata, a perennial woody and ornamental plant, was determined using ribosomal RNA-depleted total RNA (rRNA-depleted totRNA) sequencing. Five viruses  were detected and prevalence-surveyed in Hainan province, China. The genomes of polyscias capillovirus 1 (PCaV-1) and polyscias citrivirus 1 (PCiV-1) of family Betaflexiviridae were completed, and the genomes of polyscias crinivirus 1 (PCrV-1) of Closteroviridae were nearly completed lacking the 5′ and 3′ termini. PCaV-1 shares 68% genome nucleotide (nt) identity and 66% replicase (Rep) amino acid (aa) identity with homologues in apple stem grooving virus (ASGV). PCiV-1 shares 65% genome nt identity and 64% Rep aa identity with homologs in citrus leaf blotch virus (CLBV). Meeting the species demarcation criteria, PCaV-1 and PCiV-1 were considered to be new species in genera Capillovirus and Citrivirus, respectively. PCrV-1 shares high genome nt identity (62%), heat shock protein 70-like protein (HSP70h) and RNA-dependent RNA polymerase (RdRp) aa identity (78–80%) with homologues in tomato chlorosis virus (ToCV). We tentatively consider PCrV-1 to be an unclassified member of the Crinivirus genus. PoMV, PCaV-1, PCiV-1, and PCrV-1 are the prevalent viruses with >73% occurrence in the Xinglong Tropical Botanical Garden, Hainan, China.The viromic profile of  High-throughput sequencing (HTS) for viral discovery has been successfully applied to various plants, including economically important crops, perennial fruit trees, vegetables, and, more recently, wild plants ,4,5,6,7.Polyscias balfouriana, also known as Aralia balfouriana or balfour aralia, is a perennial woody plant usually cultivated as an ornamental species in gardens or indoors. This plant is attractive and highly valued for its beautiful foliage. The main cultivar is P. balfouriana Marginata, imported in 2001 from untraceable origin, and now with a small number of plantings in Hainan, China. So far, viral studies of the P. balfouriana are scarce, with only one badnavirus  being reported from the Polyscias fruticosa L.  [P. balfouriana Marginata sample  identity with different viral proteins. Similarly, 32 contigs with lengths ranging from 288 to 1343 nt hit the tomato chlorosis virus (ToCV) of the Crinivirus genus in the Closteroviridae family by sharing 32.1% to 85.9% aa identity. We thus preliminarily predicted the presence of multiple new viruses from the Betaflexiviridae and Closteroviridae families in the balfour aralia sample. aralia) . In a via sample A was cold length . We obtaP. balfouriana, we conducted rRNA-depleted totRNA sequencing to identify novel viruses, molecularly characterise them, and assess their occurrence in Hainan province, China.On the basis of dsRNA sequencing results, to further determine the virome of P. balfouriana sample showing no viral symptoms compared to the healthy plants was collected from Xinglong Tropical Botanical Garden, Hainan province, China, in 2019 , was used for host-derived read elimination using HISAT v2.1.0 [−4.After the adapter trimming and filtering of raw reads, clean reads obtained from dsRNA sequencing were de novo assembled into contigs using IDBA-UD assembler v1.1 . For rRNT v2.1.0  to reducT v2.1.0 , and annT v2.1.0  against T v2.1.0  with a c® One-Step gDNA Removal and cDNA Synthesis SuperMix  and SuperStar Max DNA Polymerase . The full genomes of the two new viruses of Betaflexiviridae were obtained with 5′ and 3′ untranslated regions (UTRs) completed, whereas the genome of the novel crinivirus of Closteroviridae lacked the 5′ and 3′ termini. The LD-oligo(dT) primer (5′-TGTGTTGGGTGTGTTTGGTTTTTTTTTTTTTTT-3′) was used to synthesise the cDNAs of RNA viruses with poly(A) tails, and the dt-rev primer (5′-TGTGTTGGGTGTGTTTGG-3′) paired with gene-specific primers was used to complete the 3′-terminal sequences. The 5′-terminal sequences were amplified using a SMARTer RACE 5′/3′ Kit  with several pairs of specific primes designed on the basis of the generated viral contigs  using Trhttps://www.ncbi.nlm.nih.gov/orffinder/ accessed on 4 October 2021), and conserved domains and motifs were identified by searching the Conserved Domain Database . Pairwise comparisons were performed on the basis of CLUSTALW multiple sequence alignments, and neighbour-joining (NJ) phylogenetic trees were constructed using MEGAX v10.1.8  [Open reading frames (ORFs) were predicted using the ORFfinder web tool   with 100P. balfouriana samples (three different cultivars) mainly exhibiting four different leaf morphologies , Closteroviridae (Crinivirus genus), and Caulimoviridae and Rhabdoviridae  [Badnavirus genus, the badnavirus found in this study was considered to be PoMV, which was for the first time discovered from P. balfouriana in China. One 5855 nt contig shared 60.57% aa identity (96% coverage) with the RdRp of cabbage cytorhabdovirus 1 (YP_010086794), which indicated the presence of a potential novel cytorhabdovirus in the Rhabdoviridae family. No further characterisation was conducted for the viruses of Caulimoviridae and Rhabdoviridae. Thirteen long contigs (1000 to 4873 nt) corresponded to criniviruses with good coverage, and were used to assemble the near-complete scaffolds of polyscias crinivirus RNA1 and RNA2 genomes in this study.The rRNA-depleted totRNA sequencing experiment yielded 76,048,918 raw reads  with Q20 and Q30 values greater than 93%, and 30,349,892 (41.8%) reads mapped to the host reference genome, while 42,341,138 (58.2%) unmapped reads were de novo assembled into 286,133 contigs. Similar to the preliminary dsRNA sequencing results, BLASTn and BLASTx analyses revealed that 1738 contigs corresponded to viruses belonging to the oviridae  familiesThe new capillovirus, tentatively given the name polyscias capillovirus 1 , consists of a positive-sense, single-stranded RNA (+ssRNA) genome of 6447 nt excluding the poly(A) tail with 5′ and 3′ UTRs completed H. Its coThe novel citrivirus, provisionally named polyscias citrivirus 1 , has an 8306 nt +ssRNA genome excluding the poly(A) tail, with a 74 nt 5′ UTR and a 201 nt 3′UTR I. The whBetaflexiviridae family showed that these capilloviruses were separated into two clades, and PCaV-1 clustered with ASGV separate from MuVA, CVA, and CuVA  for Rep and CP genes [Capillovirus and Citrivirus genera, respectively. Recently, several tentative novel capilloviruses have been identified from perennial woody plants, including loquat [Nandina domestica [P. balfouriana.On the basis of the accepted species demarcation molecular criteria for the family CP genes , polyscig loquat , birch [g loquat  and rubbg loquat . Additioomestica  and ornaomestica . Most caClosteroviridae family, provisionally named polyscias crinivirus 1 (PCrV-1), was partially molecularly characterised on the basis of long scaffolds assembled from rRNA-depleted totRNA sequencing lacking the 5′- and 3′-terminal sequences. Similar to other criniviruses, PCrV-1 has bipartite genomes of RNA1  and RNA2  with undefined 5′- and 3′-terminal sequences  with those of ToCV, while only the CP gene shares 71% aa identity satisfying the criteria.A close molecular relationship was found among PCrV-1, ToCV, and SSVV on the basis of the identities of encoded proteins  and phylCrinivirus , PCaV-1 and PCiV-1 , and the uncharacterised novel rhabdovirus . Only two samples had no target viruses (The most prevalent virus in  viruses . Althoug


This study aimed to identify key genes associated with the pathogenesis of nasopharyngeal carcinoma (NPC) by bioinformatics analysis.  Datasets (GSE13597 and GSE34573) were screened and downloaded from the comprehensive gene expression database (GEO). GEO2R online tool was adopted to analyze microarray data GSE13597 and GSE34573 related to NPC. Volcano plot was generated using Bioconductor in R software. “Pheatmap” was used to draw heatmaps based on the top 10 regulated genes of GSE13597 and GSE34573. GO and KEGG analyses were conducted via online tool DAVID. We uploaded the DEGs of NPC to STRING software and then used Cytoscape software to draw PPI network of DEGs.  216 DEGs were obtained in GSE13597 between patient and control group (111 up-regulated DEGs and 105 down-regulated DEGs). 1101 DEGs were obtained in GSE34573 (470 up-regulated DEGs and 641 down-regulated DEGs). 63 common differential genes were screened named co-DEGs in the two datasets. These DEGs were mainly associated with defense response to bacterium, cell-matrix adhesion, chemokine-mediated signaling pathway, tissue homeostasis, humoral immune response, cilium movement, cilium organization, cilium assembly, and epithelial cilium movement. KEGG pathway enrichment analysis showed that DEGs were mainly involved in viral protein interaction with cytokine and cytokine receptor, salivary secretion, p53 signaling pathway, IL-17 signaling pathway, cell cycle, PI3K-Akt signaling pathway, and ECM-receptor interaction. We identified seven hub genes, including FN1, MMP-10, MUC1, KIF23, CDK1, MUC5B, and MUC5AC.  Seven hub genes, including FN1, MMP-10, MUC1, KIF23, CDK1, MUC5B, and MUC5AC, might be therapeutic potential biomarkers of NPC. Nasopharyngeal carcinoma (NPC) is a kind of tumor characterized by high malignancy, easy metastasis with high incidence in southern China . ScientiAs an efficient and large-scale bioinformation acquisition technology, gene chip can detect and analyze differentially expressed genes (DEGs) between patients and normal humans , 5. The Present study used bioinformatics analysis to screen the related DEGs from NPC and normal nasopharyngeal tissues. Then, we conducted cluster and functional enrichment analysis of DEGs. The protein-protein interaction (PPI) network was found to further screen the hub genes. Finally, seven hub genes were obtained via intersection of two datasets, which can provide potential biomarkers for NPC for follow-up research.Homo sapiens” in the search box of the geo database website (https://www.ncbi.nlm.nih.gov/geo/) and selected keyword for retrieval. According to the search results, we obtained gene chip data GSE13597 and GSE34573. GSE13597 is based on GPL570 platform (Affymetrix Human Genome U133 Plus 2.0 Array), and GSE34573 is based on GPL76 platform (Affymetrix Human Genome U133A Array). GSE13597 consisted of 25 NPC specimens and 3 normal specimens, while GSE34573 included 3 NPC specimens and 3 normal specimens.We first entered the keywords “nasopharyngeal carcinoma” and “https://bioconductor.org/biocLite.R). The “pheatmap” package in the R statistical software was used to draw heatmaps based on the top 10 regulated genes of GSE13597 and GSE34573.Firstly, we used GEO2R tool to analyze microarray data GSE13597 and GSE34573 related to nasopharyngeal carcinoma. Volcano plot was generated using Bioconductor (https://david.ncifcrf.gov/tools.jsp), and P value <0.05 was used as the critical value for screening significantly enriched genes. In this study, the main functions of DEGs were analyzed by “clusterprofiler” package in the R software.GO and KEGG analyses were conducted via online tool DAVID (https://string-db.org/) software and applied Cytoscape software to draw PPI of DEGs.To study molecular mechanism of nasopharyngeal carcinoma cell development and pathogenesis, we uploaded the DEGs of nasopharyngeal carcinoma to STRING  . 1101 DEThe DEGs of GSE13597 were associated with antimicrobial humoral response, defense response to bacterium, cell-matrix adhesion, tissue homeostasis, negative regulation of response to wounding, anatomical structure homeostasis, and humoral immune response. GO analysis revealed the DEGs of GSE34573 associated with cilium movement, microtubule-based movement, cilium organization, axoneme assembly, cilium assembly, microtubule bundle formation, and motile cilium assembly . GO analThe DEGs screened from GSE13597 relevant to NPC were related to chemokine signaling pathway, cell cycle, p53 signaling pathway, salivary secretion, complement and coagulation cascades, and IL-17 signaling pathway . KEGG paWe submitted 216 genes of GSE13597 and 299 genes of GSE34573 to STRING for PPI analysis, respectively, and identified seven hub genes, including FN1, MMP-10, MUC1, KIF23, CDK1, MUC5B, and MUC5AC Figures .Despite the progress of radiotherapy technology, distant metastasis is still the main cause of treatment failure of NPC. Finding more effective treatment to reduce distant metastasis has attracted great attention. Therefore, understanding the etiology and mechanism of NPC progression is very important to improve survival and prevent NPC. In recent years, bioinformatics technology makes it easier for researchers to detect tumor gene changes on a large scale and has been widely used in the research field of major tumors.We screened 216 and 1101 DEGs from GSE13597 and GSE34573 datasets, respectively. These DEGs were associated with chemokine-mediated signaling pathway, cell-matrix adhesion, tissue homeostasis, defense response to bacterium, humoral immune response, cilium movement, cilium organization, cilium assembly, and epithelial cilium movement. In addition, DEGs were relevant to p53 signaling pathway, salivary secretion, complement and coagulation cascades, and IL-17 signaling pathway. The functional defects of cell cycle and cell proliferation regulators participated in the development and progression of tumor cell. Based on PPI network, seven hub genes that can deeply understand the pathogenesis of NPC at the molecular level are obtained, including FN1, MMP-10, MUC1, KIF23, CDK1, MUC5B, and MUC5AC. Xi et al. selected four genes including CRIP1, KITLG, MARK1, and PGAP1 as candidate genes affecting nasopharyngeal carcinoma-based gene expression profile analysis [Fibronectin-1 (FN1) is a member of the FN family, which was confirmed to be associated with EMT, which could enhance cancer cell invasion and metastasis , 6. FN1 MUC1 is a high molecular weight mucin rich in serine, threonine, and proline enzymes. MUC1 is anomalously expressed in numerous cancers. The coding production of MUC1 gene was a tumor marker for the diagnosis of thyroid cancer , 15. MUCHoshi et al. interfered with the expression of MUC5AC by small RNA technology and found that MUC5AC knockout in vivo inhibited the proliferation and migration of pancreatic cancer . InagumaCDKs belong to serine/threonine kinases, which play a role in all stages of the cell cycle and promote the orderly proliferation of cells . AbnormaIn conclusion, seven hub genes, including FN1, MMP-10, MUC1, KIF23, CDK1, MUC5B, and MUC5AC, might be therapeutic potential biomarkers of NPC.