沉默ski基因对大鼠星形胶质细胞活化增殖及Cyclin D1表达的影响

doi:10.3969/j.issn.1006-9771.2017.09.009

Abstract

Abstract: ObjectiveTo investigate the role of ski in proliferation of astrocytes and the molecular mechanisms in rats. MethodsAstrocytes were obtained from cerebral cortex of a three-day old rat and cultured in vitro. siRNA targeted to ski and negative control sequences were prepared. The astrocytes were divided into ski-siRNA group, siRNA negative control group and untreated control group, while the specific siRNA targeting ski negative control sequences were transfected into astrocytes with Lipofectamine^® RNAiMAX Reagent. The protein levels of ski, glial fibrillary acidic protein (GFAP) and Cyclin D1 were determined with Western blotting. The proliferation of astrocytes were measured with CCK8 assay. The cell-cycle of astrocytes were analyzed with flow cytometer. ResultsThe protein level of ski (F=38.611, P<0.01), GFAP (F=7.547, P<0.05) and Cyclin D1 (F=3.901, P<0.05) reduced in ski-siRNA group, the proliferation of astrocyte was significantly inhibited since twelve hours after culture (F>30.507, P<0.01), and less cells were in S phase and more in G1/G0 phase (F>48.425, P<0.01), compared with the control groups. Conclusionski knocking down by siRNA significantly inhibits the proliferation of astrocytes, which may associate with the down-regulation of Cyclin D1 expression.

Key words: ski, small interfering RNA, proliferation, astrocyte, rats

CLC Number:

R338.2

ZHAO Xin, GUO Yong-qiang, KOU Jiang-li, DING Ning, ZHOU Kai-sheng, NAN Wei, ZHANG Hai-hong. Effects of Knocking Down ski on Proliferation of Astrocytes and Expression of Cyclin D1 in Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2017, 23(9): 1032-1036.

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Effects of Knocking Down ski on Proliferation of Astrocytes and Expression of Cyclin D1 in Rats

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