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目的:采用RNA干扰(RNA interference,RNAi)技术沉默胃癌细胞核因子-kappa B(nuclear factor-κB,NF-κB)亚单位p65后,观察其对细胞端粒酶活性的影响。方法:采用p65小干扰RNA(small interfering RNA,siRNA)转染胃癌细胞后,分别采用western blot和凝胶迟滞电泳检测胃癌细胞p65蛋白水平和NF-κB活性,采用MTT检测胃癌细胞增殖,采用TRAP-ELISA方法检测对胃癌细胞端粒酶活性。结果:转染组细胞p65蛋白和NF-κB活性明显被抑制,且成浓度依赖性。转染组胃癌细胞增殖明显受到抑制,呈浓度依赖性。TRAP-ELISA检测显示,转染组胃癌细胞端粒酶活性明显被抑制,且呈浓度依赖性。结论:针对NF-κB重要亚单位的p65 siRNA可抑制胃癌细胞端粒酶活性。
OBJECTIVE: To observe the effect of nuclear factor-kappa B (NF-κB) subunit p65 on telomerase activity in gastric cancer cells by RNA interference (RNAi) technique. Methods: After p65 siRNA was transfected into gastric cancer cells, p65 protein and NF-κB activity in gastric cancer cells were detected by western blot and gel electrophoresis. The proliferation of gastric cancer cells was detected by MTT assay. TRAP -ELISA method for detecting telomerase activity in gastric cancer cells. Results: The p65 protein and NF-κB activity in transfected cells were significantly inhibited and in a concentration-dependent manner. The proliferation of gastric cancer cells in transfection group was significantly inhibited in a concentration-dependent manner. TRAP-ELISA assay showed that telomerase activity was significantly inhibited in a dose-dependent manner in the transfected gastric cancer cells. CONCLUSION: p65 siRNA targeting important NF-κB subunits inhibits telomerase activity in gastric cancer cells.