RNAi-mediated knockdown of INHBB increases apoptosis and inhibits steroidogenesis in mouse granulosa cells.
作者： Mohamed M’BAYE , Guohua HUA , Hamid Ali KHAN , Liguo YANG
DOI: 10.1262/JRD.2014-158
关键词:
摘要: Inhibins are members of the TGFβ superfamily and act as suppressors follicle stimulating hormone (FSH) secretion from pituitary glands via a negative feedback mechanism to regulate folliculogenesis. In this study, INHBB gene was knocked down by three RNAi-Ready pSIREN-RetroQ-ZsGreen vector- mediated recombinant plasmids explore effects silencing on granulosa cell (GC) cycle, apoptosis steroid production in vitro. Quantitative real-time polymerase chain reaction, Western blot, flow cytometry ELISA were performed evaluate role mouse GC The results showed that relative mRNA protein expression GCs can be significantly reduced RNAi with pshRNA-B1, pshRNA-B2 pshRNA-B3 plasmids, having best knockdown efficiency. Downregulation arrests cells G1 phase cycle increases rate GCs. This further confirmed downregulation expressions Cyclin D1, E Bcl2, while Bax upregulated. addition, specific markedly decreased concentration estradiol progesterone, which validated decrease levels CYP19A1 CYP11A1. These findings suggest inhibin βB is important regulation progression cells. Furthermore, subunit has biosynthesis. Evidence accumulating support concept physiologically essential for early folliculogenesis mouse.
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