【摘要】 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。更多还原

【Abstract】 Objective To investigate the effect of RNA interference targeting COX-2 on cellular proliferation and apoptosis of gastric cancer cell line SGC-7901,and provide a new pathway for gene ther- apy of gastric cancer and its recurrence and metastasis.Methods Six pieces of COX-2 small interference RNA(siRNA)and one negative control siRNA were designed and synthesized chemically,and gastric cancer cell line SGC7901 was transfecteded with them mediated by TOPtranfection.The expression of COX-2 mRNA and protein ingastric cancer cell line SGC7901 was detected by using real time quantitative PCR and Western blotting methods respectively.Cellular apoptosis was detected by flow cytometry,and compared them with controls.Results Transfection of gastric cancer cell line SGC7901 with COX- 2siRNA and negative siRNA could be mediated by TOPtranfection efficiently.Compared to negative con- trol,COX-2 mRNA in gastric cancer cell line SGC7901 measured by real time quantitative PCR after transfection with COX-2(0-5)SiRNAs mediated by TOPtranfection was decreased significantly,COX-2 siRNA resulted in highest transfection efficiency of 98.3 %,others beyond to 90 % too.Protein expres- sion of COX-2 in SGC7901 cell line measured by Western blotting was inhibited by 72.0 % and 72.1% as compared with negative control and control group respectively after transfected with COX-2 siRNA. Apoptusis rate in experimental group,negative control group and control group was 22.28±3.62,1.23±0.27 and 1.03±0.14 respectively,with the difference being significant between experimental group and negative control group(t=14.214,P＜0.01) and control group(t=14.378,P＜0.01).Conclusion RNA interference targeting COX-2 mediated by TOPtranfection can inhibit gene and protein expression of COX-2 of gastric cancer cell line SGC-7901 efficiently,and promote significantly apoptosis and suppress cellular proliferation of gastric cancer cell line SGC-7901,which provides a new pathway for gene therapy of gastric cancer and its recurrence and metastasis.更多还原