论文部分内容阅读
目的探讨靶向环氧合酶-2(COX-2)的RNA干扰(RNAi)对胃癌SGC-7901细胞增生和凋亡的影响。方法设计并合成6条COX-2小分子干扰RNA(siRNA)和1条阴性对照siR- NA,用TOPtranfection介导转染胃癌SGC7901细胞,荧光定量聚合酶链反应(PCR)和Western blot检测胃癌SGC7901细胞中COX-2基因和蛋白的表达,用流式细胞仪检测胃癌细胞凋亡情况,并与对照组进行比较。结果TOPtranfection能够有效介导COX-2 siRNA和阴性对照siRNA转染胃癌SGC7901细胞；TOPtranfection介导的COX-2 siRNA有效抑制了胃癌细胞COX-2 mRNA表达,与对照组相比抑制效率90．0%以上,最高达98．3%,同时抑制胃癌SGC7901细胞COX-2蛋白表达,与阴性对照组及对照组相比,COX-2蛋白抑制率分别为72．0%和72．1%。试验组、阴性对照组及对照组胃癌细胞凋亡率分别为：22．28±3．62、1．23±0．27和1．03±0．14。试验组与阴性对照组(t= 14．214,P＜0．01)和对照组(t=14．378,P＜0．01)比较,差异均具有统计学意义,阴性对照组与对照组比较(t=1．635,P＞0．05)差异无统计学意义。结论靶向COX2的RNA干扰能有效抑制胃癌细胞中COX-2基因和蛋白的表达,同时能明显增加胃癌细胞凋亡、抑制胃癌细胞增生。
Objective To investigate the effect of COX-2 RNAi on the proliferation and apoptosis of gastric cancer cell line SGC-7901. Methods Six small interfering RNA (siRNA) and one negative control siR-NA were designed and synthesized. SGC7901 cells were transfected with TOPtranfection, and the expression of SGC7901 was detected by fluorescence quantitative polymerase chain reaction (PCR) and Western blot. The expression of COX-2 gene and protein in cells were detected by flow cytometry and compared with the control group. Results TOPtranfection could effectively transduce COX-2 siRNA and negative control siRNA into gastric cancer SGC7901 cells; TOPtranfection-mediated COX-2 siRNA effectively inhibited COX-2 mRNA expression in gastric cancer cells, compared with the control group, the inhibitory efficiency was 90.0% , Up to 98.3%, while inhibiting COX-2 protein expression in gastric cancer SGC7901 cells compared with the negative control group and the control group, COX-2 protein inhibition rates were 72.0% and 72.1%. The apoptosis rates of gastric cancer cells in experimental group, negative control group and control group were respectively 22.28 ± 3.62, 1.23 ± 0.27 and 1.03 ± 0.14. The differences between the experimental group and the negative control group (t = 14.214, P <0.01) and the control group (t = 14.378, P <0.01) were statistically significant The difference was not statistically significant (t = 1.635, P> 0.05). Conclusions RNA interference targeting COX2 can effectively inhibit the expression of COX-2 gene and protein in gastric cancer cells, and at the same time, can significantly increase the apoptosis of gastric cancer cells and inhibit the proliferation of gastric cancer cells.