2006•Zhonghua shiyan waike zazhiRequires access
RNA interference targeting COX-2 induces gastric cancer cell line SGC-7901 apoptosis
Shirong Cai
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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.