过表达钙整合素结合蛋白CIB诱导SHG44胶质瘤细胞凋亡的实验研究

目的 观察CIB基因对人胶质瘤SHG44细胞体外生长和细胞凋亡的影响.方法 构建重组质粒pcDNA3-CIB,转染人胶质瘤细胞株SHG44,MTT观察各组细胞的体外生长情况,流式细胞术(FCM)测定各组细胞的细胞周期和凋亡细胞数量,Western-blot检测CIB转染后凋亡相关蛋白的表达变化.结果 RT-PCR、Western印迹显示pcDNA3-CIB转染组CIB的mRNA及CIB蛋白表达水平明显高于对照组;与对照组细胞相比,转染CIB的SHG44-CIB组细胞牛长明显减缓(P＜0.01),SHG44-CIB组细胞G_1、S期细胞减少,G_2期细胞增多,凋亡细胞增加至19.2%;凋亡相关蛋白Bcl-2表达下调,Bax表达上调.结论 CIB在体外明显抑制SHG44细胞的生长并诱导其发生凋亡,CIB诱导的凋亡可能与Bcl-2及Bax表达的变化相关.

Induction of apoptosis of human glioma SHG44 cells by overexpression of CIB

Objective To observe the effect of CIB gene on growth and apoptosis of human brain glioma SHG44 cells in vitro.MethodHuman glioma SHG44 cells were cultured.Recombinant plasmid pcDNA3 -CIB was constructed and transfected into SHG44 cells with Lipofectamine~(2000).A stable SHG44 cell line with overexpressing of CIB was established for the observation.The cells transfected with blank vector pcDNA3 were used as control effect of CIB on SHG44 cells.The cell growth was observed by MTr method.Flow cytometry (FCM) was used to analyze the cell cycle and apoptotic cells in each group.Then western blotting was used to detect the expression of apoptosis-associated protein.Results The stable transfected cell line SHG44-CIB was confirmed by RT -PCR and Western blotting.After transfection with CIB,SHG44 cells showed significantly decreased cell proliferation compared with the control groups.The numbers of G_1 and S phase cells were decreased,whereas G_2 phase cells were increased.The apoptotic rate reached 19.2% in SHG44-CIB cells.Overexpression of CIB decreased the expression level of bcl-2 but increased bax in SHG44 -CIB cells compared with the control groups.ConclusionsOverexpression of CIB could inhibit proliferation and induce apoptosis of SHG44.CIB induced apoptosis might work through Bax/Bcl-2 pathway.