【摘要】 目的:探讨人胰岛素样生长因子1类受体(insulin-like growth factor receptor,IGF1R)的短发夹环RNA质粒(pSUPER-siRNA-IGF1R)增加人肝癌细胞株SMMC7721对丝裂霉素诱导细胞凋亡的作用及可能的机制。方法:设计并合成特异性靶向IGF1R的siRNA片段并构建pSUPER-siRNA-IGF1R表达质粒,将其转染SMMC7721细胞,通过G418筛选出稳定株。丝裂霉素处理后,通过CCK-8法检测肝癌细胞对化疗药物敏感性的影响,流式细胞术检测各组细胞凋亡指数,Western印迹检测凋亡相关基因的变化。结果:丝裂霉素诱导细胞凋亡的作用明显增强,凋亡指数明显升高(P<0.05)。实验组野生型p53蛋白,bax蛋白表达比对照组明显上调,而bcl-2蛋白表达明显下降。结论:构建的pSUPER-siRNA-IGF1R具有RNA干扰作用,并能增强丝裂霉素诱导的细胞凋亡。更多还原

【Abstract】 Objective:To investigate whether the plasmid of short hairpin RNA targeting human insulin like growth factor receptor 1(pSUPER-siRNA-IGF1R) can enhance mitomycin-induced apoptosis of human liver cancer SMMC7721 cells and the corresponding mechanism.Methods:The siRNA targeting IGF1R was designed and the pSUPER-siRNA-IGF1R plasmid was constructed and transfected into SMMC7721 cells.The stable cell lines were screened by G418.The growth curve of every group was detected by CCK8 method.After mitomycin treatment,the sensitivity of hepatoma cells to chemotherapy was monitored by CCK8 method.The apoptotic index(AI)was examined by flow cytometry.The expression of apoptosis-related proteins p53,Bax,bcl-2 was determined by Western blotting.The empty control group and positive control were set up.Results:The apoptosis induced by mitomycin were significantly enhanced in stable cell line and the AI was significantly elevated compared with other groups(P<0.05).The expression of wild type p53 and Bax protein was significantly up-regulated but the expression of bcl-2 protein was significantly down-regulated(P<0.05).Conclusion:The constructed pSUPER-siRNA-IGF1R plasmid silences RNA transcription and enhances the apoptosis of tumor cells induced by mitomycin.更多还原