MiR-106a-5p modulates apoptosis and metabonomics changes by TGF-β/Smad signaling pathway in cleft palate.
BACKGROUND
The molecular mechanisms of abnormal palatogenesis were investigated in this study. A key regulator, miR-106a-5p, and its target pathway were analyzed.


OBJECTIVES
This research is trying to clarify the underlying mechanism of the modulation of miRNA transcription during the formation of cleft palate by 7T and 9.4T NMR metabolomic platforms.


METHOD
Differentially expressed miRNAs and mRNAs were analyzed by microarray analysis and verified by qRT-PCR. The protein expression in TGFβ signaling pathways were analyzed by Western Blotting. The relationship between miR-106a-5p and TGFβ were analyzed by luciferase reporter assay. Cell apoptosis were analyzed by flow cytometer. And finally, the metabonomics were analyzed by NMR and multivariate data analysis models (MVDA).


RESULTS
The expression of miR-106a-5p increased in cleft palatal tissue and negatively correlated with the protein level of Tgfbr2. The luciferase assay further proved that the tgfbr2 was a direct target of miR-106a-5p. In another aspect, miR-106a-5p increased apoptosis level in palatal mesenchymal cells, possibly because its inhibition of TGFβ signaling pathway. Moreover, low cholesterol and choline levels with high citric acid and lipid levels were observed by 7T and 9.4T NMR metabonomic analysis, which inferred the disorder of cell membrane synthesis in cleft palate formation. Furthermore, transformation from choline to phosphatidylcholine regulated by miR-106a-5p was also disrupted, resulting in phosphatidic choline synthesis disorder and reduced cell membrane synthesis.


CONCLUSIONS
The regulatory mechanism of cleft palate was studied at transcriptional and metabolomics levels, which may provide important information in understanding the primary cause of this abnormality.
['Zhang|Wancong|W|', 'Shen|Zhiwei|Z|', 'Xing|Yue|Y|', 'Zhao|Hanxing|H|', 'Liang|Yan|Y|', 'Chen|Jiasheng|J|', 'Zhong|Xiaoping|X|', 'Shi|Lungang|L|', 'Wan|Xinhong|X|', 'Zhou|Jianda|J|', 'Tang|Shijie|S|']
D000818:Animals D017209:Apoptosis D002462:Cell Membrane D019343:Citric Acid D002972:Cleft Palate D004195:Disease Models, Animal D004622:Embryo, Mammalian D005260:Female D018507:Gene Expression Regulation, Developmental D006801:Humans D008297:Male D008563:Membrane Lipids D059630:Mesenchymal Stem Cells D055442:Metabolome D051379:Mice D035683:MicroRNAs D010159:Palate D000077294:Receptor, Transforming Growth Factor-beta Type II D015398:Signal Transduction D051899:Smad2 Protein D051900:Smad3 Protein D059467:Transcriptome D016212:Transforming Growth Factor beta D014212:Tretinoin
Cleft palate Metabolomics MicroRNAs