COX-2特异性的siRNA诱导胃癌SGC-7901细胞凋亡

siRNA targeting COX-2 induces gastric carcinoma cell SGC-7901 apoptosis

目的利用RNA干扰(RNAi)技术在体外抑制COX-2基因表达,探讨RNAi诱导胃癌SGC-7901细胞凋亡的可能途径。方法设计靶向COX-2基因的小干扰RNA(si RNA),构建重组表达质粒pTZU6+1-si RNA-COX-2,并导入SGC-7901细胞,在体外诱导RNAi,用RT-PCR、免疫细胞化学、末端标记细胞凋亡检测法(TUNEL)和Western Blot检测COX-2基因表达、细胞凋亡和凋亡相关基因的表达。结果重组质粒pTZU6+1-si RNA-COX-2导入SGC-7901细胞株后,COX-2表达明显下调,细胞出现凋亡(P<0.05),Caspase-9和Caspase-3表达明显上调。结论 RNA干扰明显抑制了靶基因COX-2的表达,并可能通过激活Caspase途径诱导SGC-7901细胞凋亡。

Objective To investigate the possible pathway of SGC-7901 cells apoptosis by utilizing RNA interference（RNAi） to inhibit expression of COX-2 in vitro.Methods Small interference RNA（siRNA） homologous to COX-2 gene were designed,pTZU6＋1-siRNA-COX-2 vector was constructed and transfected into SGC-7901 cells to induce RNAi.The changes of COX-2 gene,cell apoptosis and expression of apoptosis related genes were detected with RT-PCR,immunochemistry,TUNEL and Western blottingin.Results After transfected with pTZU6＋1-siRNA-COX-2,the level of cox-2 gene was down-regulated,SGC-7901 cell apoptosis was obvious（P〈0.05）,expressions of Caspase-9 and Caspase-3 were arised.Conclusion RNA interference down-regulate COX-2 expression,and induce SGC-7901 cell apoptosis maybe by exciting expression of Caspases.