Effects of bta-miR-101 on Proliferation,Apoptosis and Secretion of Bovine Testicular Sertoli Cells
The study aimed to explore the effects of bta-miR-101 on bovine testicular sertoli cells'proliferation,apoptosis and secretion.In this study,testicular tissues of healthy bulls(n=3)aged 3 days and 13 months were collected to construct the expression profile of Angus cattle tissues.The differences in bta-miR-101 expression were verified between the two periods.Online tools(TargetScan,miRTarBase,miRDB,and miRWalk)were used to predict the target gene of miR-101,and KOBAS was used for enrichment analysis of GO and KEGG pathways.The mimics and inhibitors of miR-101 were transfected into bovine testicular sertoli cells.Cell proliferation,apoptosis,secretion-related gene expression,and cell proliferation coefficient were detected by RT-qPCR,Western Blotting,and CCK-8 techniques.There were 26 common target genes for bta-miR-101 predicted by 4 online tools.GO and KEGG enrichment analysis showed that these genes were mainly enriched in the nucleoplasm,protein-binding,and JAK-STAT receptor signaling pathways,which were associated with cell differentiation,cell cycle,apoptosis,and other biological processes to varying degrees.The results of RT-qPCR showed that bta-miR-101 expression was significantly higher in testis tissue at birth than at sexual maturity(P＜0.05),consistent with previous laboratory sequencing results.After transfection of miR-101 mimics and inhibitors,it was found that miR-101 mimics could promote the expression of proliferation-related genes at mRNA and protein levels,significantly increase the proliferation coefficient of sertoli cells,and inhibit the expression of apoptosis genes at mRNA and protein levels(P＜0.05);miR-101 inhibitors inhibited the expression of proliferation-related genes at mRNA and protein levels and promoted the expression of apoptotic genes at mRNA and protein levels(P＜0.05).In terms of cell secretion,miR-101 had an effect on the transcription of secretion marker genes GDNF and BMP4,but had no significant effect on the secretion of androgen-binding proteins.In conclusion,miR-101 promoted proliferation and inhibited apoptosis of immature sertoli cells of the bovine testis,and had no significant effect on the secretion of androgen-binding proteins.
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