Effects of Knocking Down ski on Proliferation of Astrocytes and Expression of Cyclin D1 in Rats
10.3969/j.issn.1006-9771.2017.09.009
- VernacularTitle: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
- Author:
Xin ZHAO
;
Yongqiang GUO
;
Jiangli KOU
;
Ning DING
;
Kaisheng ZHOU
;
Wei NAN
;
Haihong ZHANG
- Keywords:
ski;
small interfering RNA;
proliferation;
astrocyte;
rats
- From:
Chinese Journal of Rehabilitation Theory and Practice
2017;23(9):1032-1036
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of ski in proliferation of astrocytes and the molecular mechanisms in rats. Methods Astro-cytes 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 spe-cific 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. Results The 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. Conclusion ski knocking down by siRNA significantly inhibits the proliferation of astro-cytes, which may associate with the down-regulation of Cyclin D1 expression.
