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小干扰RNA介导的BMPRIB基因沉默对猪颗粒细胞凋亡及类固醇生成的影响

Effect of BMPRIB gene silencing by siRNA on apoptosis and steroidogenesis of porcine granulosa cells.

作者信息

Zhao Y Y, Li X X, Wang W, Chen X, Yu P, Wang J J, Xu Y X

机构信息

College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China.

Harbin Veterinary Research Institute, Chinese Academy of Agricultural Science, Harbin, China.

出版信息

Genet Mol Res. 2014 Nov 28;13(4):9964-75. doi: 10.4238/2014.November.28.1.

Abstract

Bone morphogenetic proteins (BMPs) are the key factors in maintaining cell growth and differentiation in ovaries. BMPs initiate signaling by assembling BMP receptors and activating Smads, which in turn alter the expression of target genes. However, little is known about the effect of the deletion of the Bone morphogenetic protein receptor type IB (BMPRIB) on porcine granulosa cell (GCs). The objective of this study was to determine the effects of BMPRIB gene silencing, by small interfering RNA (siRNA), on the apoptosis and steroidogenesis of porcine GCs, and the expression of cell cycle-related and apoptosis-related genes. Results indicate that the BMPRIB siRNA caused specific inhibition of BMPRIB mRNA expression after transfection. Knockdown of the BMPRIB gene significantly inhibited porcine GCs proliferation and estradiol production, while inducing apoptosis of porcine GCs. Additionally, the declined expression of the BMPRIB gene changed the expressions of CylinD2, Cdk2, Bcl-2, and Cyp19a1. These findings provide an important role of BMPRIB in the regulation of apoptosis and steroidogenesis of porcine GCs.

摘要

骨形态发生蛋白(BMPs)是维持卵巢细胞生长和分化的关键因子。BMPs通过组装BMP受体并激活Smads来启动信号传导,进而改变靶基因的表达。然而,关于骨形态发生蛋白受体IB型(BMPRIB)缺失对猪颗粒细胞(GCs)的影响知之甚少。本研究的目的是确定通过小干扰RNA(siRNA)沉默BMPRIB基因对猪GCs凋亡、类固醇生成以及细胞周期相关基因和凋亡相关基因表达的影响。结果表明,转染后BMPRIB siRNA对BMPRIB mRNA表达产生了特异性抑制。敲低BMPRIB基因显著抑制了猪GCs的增殖和雌二醇的产生,同时诱导了猪GCs的凋亡。此外,BMPRIB基因表达的下降改变了细胞周期蛋白D2(CylinD2)、细胞周期蛋白依赖性激酶2(Cdk2)、B细胞淋巴瘤-2(Bcl-2)和细胞色素P450 19A1(Cyp19a1)的表达。这些发现揭示了BMPRIB在调节猪GCs凋亡和类固醇生成中具有重要作用。

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