热休克蛋白70羧基末端相互作用蛋白小干扰RNA对人脑胶质瘤细胞周期及增殖的影响

Effect of small interfering carboxyl terminus of Hsc70-interacting protein on cell cycle and proliferation of human brain glioma cells

目的 观察热休克蛋白70羧基末端相互作用蛋白（CHIP）小干扰RNA（siRNA）对人脑胶质瘤细胞周期及增殖的影响.方法 运用Control siRNA、CHIP siRNA分别转染原代培养人脑胶质瘤细胞,Western blot检测CHIP蛋白表达,流式细胞仪检测CHIP siRNA对原代培养人脑胶质瘤细胞周期的影响,细胞计数试剂盒（CCK-8）细胞增殖实验检测CHIP siRNA对原代培养人脑胶质瘤细胞增殖的影响.结果 与Control siRNA组比较CHIP siRNA组原代培养人脑胶质瘤细胞CHIP蛋白表达量降低53.3％,CHIP siRNA组原代培养人脑胶质瘤细胞增殖细胞数增加,并且G1期细胞数所占比例减少14.1％,S期细胞数所占比例增加13.7％ （P ＜0.05）.结论 CHIP siRNA靶向干扰了CHIP的表达,CHIP作为肿瘤抑癌因子抑制脑胶质瘤细胞的增殖,并能使细胞阻滞在G1期.

Objective To investigate the effect of carboxyl terminus of Hsc70-interacting protein （CHIP） small interfering RNA （siRNA） on cell cycle and proliferation of human brain glioma cells.Methods Control siRNA and CHIP siRNA were used respectively to transfect primary cultured human brain glioma cells.The expression levels of CHIP in primary cultured human brain glioma cells were detected after CHIP knockdown by using Western blotting.The influence of CHIP knockdown on the cell cycle of primary cultured human brain glioma cells was measured by flow cytometry.The influence of CHIP knockdown on the proliferation of primary cultured human brain glioma cells was tested by using cell counting kit-8 （CCK-8）assay.Results As compared with control siRNA group,the expression of CHIP was decreased by 53.3％ in CHIP siRNA group.CHIP knockdown could drastically increase the ability of proliferation in primary cultured human brain glioma cells,increase the S phase ratio of cell cycle by 13.7％ and decrease the G1 phase ratio of cell cycle by 14.1％ （both P ＜ 0.05）.Conclusion CHIP-targeted siRNA interferes with the expression levels of CHIP.CHIP acts as a tumor suppressor that suppresses the proliferation of glioma cells and arrests the cells in G1 phase.