miR-139对猪未成熟支持细胞增殖和凋亡调控的初步研究

The role of miR-139 in regulating proliferation and apoptosis of porcine immature sertoli cells

支持细胞的增殖力决定性成熟后睾丸支持细胞的数量,进而影响公猪的精子发育、成熟及精液品质。microRNA通过结合靶基因的3′-UTR,广泛参与调控支持细胞增殖、凋亡和分化等多种生物学过程。为了研究miR-139在猪未成熟支持细胞中的作用,试验将miR-139过表达试剂、抑制表达试剂及真核翻译起始因子(EIF4G2)的干扰试剂分别转染到猪的未成熟支持细胞中,采用实时荧光定量及流式细胞术等方法对其进行研究。结果显示:过表达miR-139,细胞周期进程加快,增殖能力显著增强,而细胞凋亡率则显著降低;抑制表达miR-139后,结果与之相反。干扰EIF4G2基因后,细胞周期进程加快,且促进了细胞增殖而抑制其凋亡,与过表达miR-139的结果相一致。结果说明miR-139促进猪未成熟支持细胞增殖而抑制其凋亡,且其作用机制可能是通过靶向EIF4G2基因实现。该研究为进一步探讨支持细胞增殖对精子发生的作用提供理论依据。

The proliferative activity of sertoli cells determines the number of such cells in the testis after sexual maturation,which in turn affects sperm development,maturation and semen quality in boars.microRNAs are widely involved in regulating various biological processes such as proliferation,apoptosis and differentiation of sertoli cells by binding to the 3′-UTR of target genes.To investigate the role of miR-139 in porcine immature sertoli cells,miR-139 overexpression reagent,inhibitory expression reagent and EIF4G2 gene interference reagent were transfected into the cells.the real-time fluorescence quantification,flow cytometry and other experiments were performed to assay these roles.The results showed that the overexpression of miR-139 significantly accelerated the progression of the cell cycle,significantly increased their proliferation and decreased the rate of their apoptosis;and the opposite result was observed after inhibition of the miR-139 expression.Inhibition of the expression of the EIF4G2 gene promoted the cell cycle of entry into the S phase and enhanced cell proliferation while inhibi⁃ting their apoptosis,which was consistent with the results of miR-139 overexpression.These results suggested that miR-139 promoted prolif⁃eration and inhibited apoptosis in porcine immature sertoli cells,and this mechanism of action mgiht be achieved by targeting the EIF4G2 gene.The present study provided a theoretical basis for further research on the role of sertoli cell proliferation in spermatogenesis.