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目的研究靶向HER2/neu基因的siRNA对膀胱癌BIU-87细胞增殖和凋亡的影响。方法化学合成的靶向HER2/neu基因siRNA在脂质体的介导下转染BIU-87细胞,实验分为HER2/neu siRNA组、空脂质体组、阴性siRNA序列组,以未转染BIU-87细胞为空白对照组。利用CCK-8法评价HER2/neu基因siRNA对BIU-87细胞体外生长的抑制作用,流式细胞术检测细胞凋亡情况,通过逆转录聚合酶链反应(RT-PCR)和Western blot法检测转染前后细胞中HER2/neu mRNA和蛋白表达水平的变化。结果转染HER2/neu siRNA后BIU-87细胞的存活率显著下降,从(82.37±0.90)%降低到(56.76±1.70)%,差异有统计学意义(P<0.05);同时能诱导细胞凋亡,HER2/neu siRNA组凋亡率达(45.60±0.70)%,与空白对照组、空脂质体组、阴性siRNA序列组比较差异有统计学意义(P<0.05);靶向HER2/neu基因siRNA能显著降低BIU-87细胞中HER2/neu mRNA和蛋白的表达(P<0.05)。结论化学合成的靶向HER2/neu基因siRNA能有效抑制HER2/neu基因在BIU-87细胞中的表达,进而能有效抑制BIU-87细胞的增殖和诱导凋亡的作用。
Objective To study the effect of siRNA targeting HER2 / neu gene on the proliferation and apoptosis of bladder cancer BIU-87 cells. Methods Chemically synthesized siRNA targeting HER2 / neu gene was transfected into BIU-87 cells mediated by liposomes. The experiment was divided into HER2 / neu siRNA group, empty liposome group, negative siRNA sequence group, untransfected BIU-87 cells as a blank control group. The inhibitory effect of HER2 / neu gene siRNA on the growth of BIU-87 cells in vitro was evaluated by CCK-8 assay. The apoptosis of BIU-87 cells was evaluated by flow cytometry. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot Changes of HER2 / neu mRNA and protein expression in the cells before and after dyeing. Results The survival rate of BIU-87 cells after transfection with HER2 / neu siRNA was significantly decreased from (82.37 ± 0.90)% to (56.76 ± 1.70)%, the difference was statistically significant (P <0.05) The apoptosis rate of HER2 / neu siRNA group was (45.60 ± 0.70)%, which was significantly lower than that of blank control group, empty liposome group and negative siRNA sequence group (P <0.05) Gene siRNA can significantly reduce HER2 / neu mRNA and protein expression in BIU-87 cells (P <0.05). Conclusion Chemically synthesized HER2 / neu gene siRNA can effectively inhibit the expression of HER2 / neu gene in BIU-87 cells, which can effectively inhibit the proliferation and induce apoptosis of BIU-87 cells.