TGF-β调节仔猪睾丸支持细胞增殖的机制

doi:10.11843/j.issn.0366-6964.2018.04.006

Abstract

Abstract:

The objective of the study was to investigate the effect of TGF-β on the expression of proliferation-related proteins and genes of boar Sertoli cells, and reveal the mechanism of TGF-β regulating proliferation of boar Sertoli cells. In this study, TGF-β and TGF-β/Smad pathway inhibitor LY2109761 was used to stimulate cultured immature boar Sertoli cells. Cell viability and cell cycle were detected by CCK-8 kit and flow cytometry, respectively. The phosphorylation levels of Smad3 were detected by Western blotting. The expressions of c-Myc mRNA, Skp2 mRNA and ssc-miRNA-24 were detected by real-time quantitative PCR (RT-qPCR). Furthermore, ssc-miRNA-24 mimics and inhibitor were transfected into Sertoli cells, and then Sertoli cells were treated with TGF-β, following detected the expression of related genes and proteins. The results showed that TGF-β influenced cell viability in a dose-dependent manner. 180 pg·mL^-1 TGF-β increased cell proliferation index (PI)and S-phase cell fraction (SPF), inhibited the phosphorylation of Smad3 in a time-dependent (0-24 h) manner, increased the abundance of c-Myc mRNA, Skp2 mRNA, ssc-miRNA-24. 10 μmol·L^-1 LY2109761 promoted TGF-β-induced cell viability, increased cell proliferation index and S-phase cell fraction. In addition, ssc-miRNA-24 mimics decreased phosphorylation level of Smad3, promoted the expression of c-Myc and Skp2 mRNA. However, ssc-miRNA-24 inhibitor had an opposite effect on these parameters. The above results revealed that TGF-β decreased phosphorylation level of Smad3 by increasing the expression of ssc-miRNA-24, and enhanced the expression of c-Myc and Skp2 mRNA to promote the proliferation of boar Sertoli cells.

CLC Number:

S828.2

HUANG Sha, SUN Si, DENG Jie, HU Yu, LIAN Yu, WANG Xian-zhong. Mechanism of TGF-Beta Regulating Proliferation of Boar Sertoli Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(4): 701-708.

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Mechanism of TGF-Beta Regulating Proliferation of Boar Sertoli Cells

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