EMMPRIN基因表达对胶质瘤U251细胞增殖、凋亡的影响

摘要

目的 探讨特异性小分子干扰(small interference,siRNA)抑制人脑胶质瘤U251细胞株内细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,EMMPRIN)基因的表达后,对U251细胞株增殖、凋亡的影响.方法 构建EMMPRIN基因小干扰RNA,转染人胶质瘤U251细胞株,运用半定量RT-PCR及Western blot的方法验证转染组及对照组的胶质瘤细胞EMMPRIN基因mRNA及蛋白的表达水平,通过MTT法观察转染48 h后对U251细胞增殖的变化,流式细胞术观察转染48 h后U251细胞周期及凋亡的改变.结果 EMMPRIN基因的特异性siRNA转染U251细胞后,EMMPRIN基因的mRNA和蛋白表达水平明显降低,细胞增殖明显受到抑制,细胞凋亡增加.结论 EMMPRIN基因的特异性siRNA能抑制胶质瘤U251细胞株的增殖,促进细胞凋亡.%Objective To investigate the effect of downregulating EMMPRIN gene expression by RNA interference on proliferation and apoptosis of glioblastm U251 cells. Methods EMMPRIN gene of small interference RNA was constructed. Human glioma U251 cell line was transfected. EMMPRIN mRNA gene and protein expression levels were validated using the semi quantitative RT -PCR and Western blotting in transfection group and control group . 48 hours after transfection, U251 cell proliferation changes were observed using the method of MTT and the change of cell cycle and apoptosis in U251 cells was observed by flow cytometry. Results After the EMMPRIN gene specific siRNA was transfected with U251 cells, the expression level of EMMPRIN gene mRNA and protein significantly decreased, cell proliferation was inhibited, and apoptosis increased. Conclusions Downregulating the expression of EMMPRIN gene by siRNA can inhibit prolifetation and induce apoptosis in human glioblastoma U251 cells. Therefore, a siRNA - targeted EMMPRIN gene strategy is expected to be a protentially ideal target for gene therapy of gliomas.