RNA干扰下调肿瘤相关钙信号传导蛋白-2基因表达对胃癌细胞黏附、迁移和侵袭的影响

目的 观察肿瘤相关钙信号传导蛋白-2(TROP-2)基因小干扰RNA(siRNA)对胃癌细胞生物学行为的影响.方法 培养人胃癌MGC-803、HGC-27及BGC-823细胞株,以荧光实时定量聚合酶链反应(PCR)检测TROP-2基因mRNA表达;筛选出TROP-2表达最高者.采用TROP-2基因小干扰RNA转染胃癌细胞株,分别以荧光实时定量PCR和免疫荧光方法观察TROP-2基因mRNA和蛋白水平,然后以计数法检测细胞黏附,以划痕法观察癌细胞迁移、以Boyden方法检测癌细胞侵袭力.结果 荧光实时定量PCR方法显示,3株胃癌细胞中,TROP-2均有不同程度的表达,以胃癌BGC-823细胞最高;以TROP-2 siRNA转染胃癌BGC-823细胞后,癌细胞TROP-2基因mRNA和蛋白明显下降,且呈浓度依赖性;细胞黏附结果显示,转染组细胞黏附数量明显下降;划痕试验和Boyden试验结果显示,细胞迁移和侵袭力明显下降.结论 TROP-2基囚在胃癌细胞黏附、迁移和侵袭中发挥着重要作用;以siRNA转染胃癌细胞,可抑制胃癌细胞黏附、迁移和侵袭能力.

Effects of down-regulation of tmnor-associated calcium signal transducer-2 gene expression by RNA interference on adhesion, migration and invasion of gastric cancer cells

Objective To study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA (siRNA) on adhesion, migration, and invasion of human gastric cancer cells.Methods Real time polymerase chain reaction (PCR) was used to detect the TROP-2 mRNA expression of human gastric cancer cell lines MGC-803, HC, C-27 and BGC-823. The cells with highest expression of TROP-2 were transfected with different doses of TROP-2 siRNA. The expression of TROP-2 mRNA and protein was detected by real-time quantitative PCR and immumoflureseence method. Cell adhesion, migration,and invasion were exmined by hoyden chamber, respectively. Results Cell line BGC-823 showed the highest elevation of TROP-2 mRNA among three gastric cancer cell lines. Real-time quantitative PCR and immumoflurescence method revealed that the expression of TROP-2 mRNA and protein was reduced in a time- and dose-dependent manner ( r = 0. 935 ; r = 0. 922). The ability of adhesion, migration and invasion of BGC-823 cells treated with TROP-2 siRNA was decreased as compared with control group (P <0. 01 ). Conclusion TROP-2 gene might play an important role in adhesion, migration, and invasion of human gastric cancer cells, siRNA targeted TROP-2 could effectively inhibit adhesion, migration, and invasion of human gastric cancer cells.