Effect of Knocking Down ski on Migration of Astrocytes in Rats

ZHAO Xin; WANG Xing-wen; KOU Jiang-li; LI Zhong-hao; GUO Yong-qiang; WU Ya-min; ZHANG Hai-hong

doi:10.3969/j.issn.1006-9771.2017.08.008

Abstract：Objective To investigate the effect of ski gene in migration process of astrocytes in rats. Methods Astrocytes were obtained from rats' cerebral cortex and cultured in vitro. siRNA targeting ski gene and negative control sequences were prepared. The ski-siRNA group, siRNA negative control group and untreated group were set in this experiment. The specific siRNA targeting ski gene was transfected into astrocytes with Lipofectamine?RNAiMAX Reagent. Then the ski protein levels were determined with Western blotting. After transfec-tion, the changes in migration of astrocytes were measured with wound scratch assay and Transwell migration assay. Results Western blot-ting showed that the expression of ski protein was significantly lower in the ski-siRNA group than in the siRNA negative control group and untreated group (F=132.957, P<0.001). Transwell migration assay showed that the number of astrocytes crossing through chambers was less in the ski-siRNA group than in the siRNA negative control group and untreated group (F>47.197, P<0.05). Wound scratch assay showed that the wound healing rate was lower in the ski-siRNA group than in the control group one, two, three, four and five days after transfection (F>69.187, P<0.001). Conclusion Ski knocked down by siRNA could inhibit the migration ability of astrocytes. It is a reminding that ski may take part in the migration process of astrocytes, and moreover, ski may play an important role in the formation of glial scar.