miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene

RAN Mao-liang; WENG Bo; CAO Rong; PENG Fu-zhi; LUO Hui; GAO Hu; CHEN Bin

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miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene

机译：miR-34c通过靶向SMAD7基因抑制未成熟猪Sertoli细胞的增殖并增强其凋亡

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MicroRNAs (miRNAs) are implicated in swine spermatogenesis via their regulations of cell proliferation, apoptosis, and differentiation. Recent studies indicated that miR-34c is indispensable in the late steps of spermatogenesis. However, whether miR-34c plays similar important roles in immature porcine Sertoli cells remain unknown. In the present study, we conducted two experiments using a completely randomised design to study the function roles of miR-34c. The results from experiment Ⅰ demonstrated that the relative expression level of miR-34c in swine testicular tissues increased (P=0.0017) quadratically with increasing age, while the relative expression level of SMAD family member 7 (SMAD7) decreased (P=0.0009) with curve. Furthermore, miR-34c expression levels showed a significant negative correlation (P=0.013) with SMAD7 gene expression levels. The results from experiment Ⅱ indicated that miR-34c directly targets the SMAD7 gene using a luciferase reporter assay, and suppresses (P＜0.05) SMAD7 mRNA and protein expressions in immature porcine Sertoli cells. Overexpression of miR-34c inhibited (P＜0.05) proliferation and enhanced (P＜0.05) apoptosis in the immature porcine Sertoli cells, which was supported by the results from the Cell Counting Kit-8 (CCK-8) assay, the 5-Ethynyl-2′-deoxyuridine (EdU) assay, and the Annexin V-FITC/PI staining assay. Furthermore, knockdown of SMAD7 via small interfering RNA (siRNA) gave a similar result. It is concluded that miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene.

机译：MicroRNA（miRNA）通过细胞增殖，凋亡和分化的调控参与猪的精子发生。最近的研究表明，在精子发生的后期，miR-34c是必不可少的。然而，miR-34c在未成熟的猪支持细胞中是否起着类似的重要作用仍是未知的。在本研究中，我们使用完全随机的设计进行了两个实验，以研究miR-34c的功能作用。实验Ⅰ的结果表明，miR-34c在猪睾丸组织中的相对表达水平随着年龄的增长呈二次增加（P = 0.0017），而SMAD家族成员7（SMAD7）的相对表达水平则随着年龄的增长而降低（P = 0.0009）。曲线。此外，miR-34c表达水平与SMAD7基因表达水平呈显着负相关（P = 0.013）。实验Ⅱ的结果表明，miR-34c通过荧光素酶报告基因直接靶向SMAD7基因，并抑制（P ＜0.05）未成熟猪睾丸支持细胞中SMAD7的mRNA和蛋白表达。 miR-34c的过表达抑制未成熟猪Sertoli细胞的（P <0.05）增殖并增强（P <0.05）的细胞凋亡，这由细胞计数试剂盒8（CCK-8）测定结果支持，5-乙炔基2'-脱氧尿苷（EdU）测定和膜联蛋白V-FITC / PI染色测定。此外，通过小干扰RNA（siRNA）敲低SMAD7的结果相似。结论是，miR-34c通过靶向SMAD7基因抑制未成熟猪Sertoli细胞的增殖并增强其凋亡。

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《农业科学学报（英文版）》 |2019年第2期|449-459|共11页
作者
RAN Mao-liang; WENG Bo; CAO Rong; PENG Fu-zhi; LUO Hui; GAO Hu; CHEN Bin;
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College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China;

College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China;

College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China;

College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China;

College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China;

College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China;

College of Animal Science and Technology/Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, P.R.China;

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miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene

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