MACC1表达沉默对胶质瘤细胞U87增殖凋亡的影响

The effects of silencing MACC1 gene by RNAi on proliferation and apoptosis of U87 human brain glioma cells

目的探讨MACC1基因表达沉默对脑胶质瘤细胞U87增殖凋亡的影响。方法应用RT-PCR和Western blot检测55例不同病理级别胶质瘤标本MACC1表达;转染MACC1siRNA到U87细胞沉默MACC1基因表达,Western blot检测MACC1,Bcl-2/Bax,caspase-3,c-Met蛋白的表达;MTT法检测细胞生长增殖能力;流式细胞术检测细胞周期及凋亡情况。结果MACC1基因在胶质瘤标本组织表达随病理级别升高而增多;MACC1表达沉默后U87生长增殖能力明显下降,细胞周期阻滞于G0/G1期,细胞凋亡显著增加,MACC1蛋白表达明显下降,Bcl-2/Bax比值降低,caspase-3表达上调,c-Met表达下降。结论下调MACC1表达可促进胶质瘤细胞凋亡,抑制肿瘤细胞增殖,MACC1可能成为胶质瘤基因治疗的新靶点。

Objective To investigate the effects of silencing MACC1 gene by RNAi on the proliferation and apoptosis of U87 human brain glioma cells. Methods The expression of MACC 1 in different pathological grade gliomas of 55 patients was detected by RT-PCR and Western blot. MACClsiRNA was transfected into U87 cells to silence MACC1 gene expression, the expressions of c-Met, Bcl-2/Bax, Caspase-3, and MACC1 protein were detected by Western blot. Cell proliferation was detected by MT1~, Cell cycle and apoptosis were measured by flow cytometry. Results The expression level of MACC 1 gene in U87 cells was increased as the increased pathological grade of glioma specimens. After MACC 1 silencing, the proliferation rate of U87 was decreased significantly, cell cycle arrested in G0/G1 phase, and the cell apoptosis was significantly increased, MACC1 protein expression level was significantly decreased, Bcl-2/Bax ratio was decreased, the expression levels of caspase-3 and c-Met were decreased. Conclusion MACC1 might be involved in the pathogenesis of glioma, and a new target for gene therapy of glioma.