Ran Mao-liang;Weng Bo;Cao Rong;Peng Fu-zhi;Luo Hui;Gao Hu;Chen Bin

Hunan Agr Univ, Hunan Prov Key Lab Genet improvement Domest Anim, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China

immature porcine Sertoli cell;cell proliferation;apoptosis;SMAD7;miR-34c

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.