RNAi对胃癌SGC-7901细胞侵袭及迁移能力的影响

The influence of RNAi on invasion and migration of gastric cancer cell line SGC-7901

目的运用RNA干扰(RNAi)技术抑制HER2基因的表达,探讨其对胃癌SGC-7901细胞侵袭及迁移能力的影响。方法设计、构建RNAi片段靶向抑制HER2高表达的胃癌细胞株(HER2-RNAi组),以转染阴性对照系列(阴性对照组)及未转染的SGC-7901细胞(空白对照组)为对照组。利用RT-PCR和Western blot法检测各组胃癌SGC-7901细胞中HER2的表达情况;采用Transwell小室模型及划痕实验检测RNAi对胃癌细胞体外的侵袭和迁移能力的影响。结果 RNAi片段转染后SGC-7901细胞的HER2mRNA和蛋白表达水平明显下降(P<0.05);侵袭实验结果显示,HER2-RNAi组的穿膜细胞数[(31.5±3.8)个/视野]显著低于未转染组[(103.6±4.5)个/视野];迁移实验结果显示,HER2-RNAi组的穿膜细胞数[(63.6±5.1)个/视野]显著低于未转染组[(193.5±6.2)个/视野],各组间比较差异有统计学意义(P<0.05)。结论 RNAi沉默HER2基因可以抑制胃癌SGC-7901细胞的侵袭和迁移能力,提示RNAi基因沉默技术有可能成为进展期胃癌治疗的新方法。

Objective To investigate the effect of RNA interference（RNAi） on gastric cancer cell line SGC-7901 invasion and migration by inhibiting HER2 expression. Methods The RNAi segments targeting HER2 were designed and transfected into cell lines with high level of HER2 （HER2-RNAi group） ,SGC-7901 cells transfected with nagetive control series（the nagetive control group） and vacant SGC-7901 cells（the blank control group） were used as controls. The mRNA and protein expression of HER2 were detected by real-time PCR and Western blot. The invasive and migration ability of transfected tumor cells by Transwell assay and scratch assay. Results The expression of HER2 Was significantly reduced after the transfection（P〈0.05）. In invasion test,the number of cells crossing through chambers in HER2-RNAi group[ （31.5 ±3.8）/view3 was significantly lower than that in the control group[ （103.6± 4. 5）/view3 （P〈0.05）; in migration test, the number of cells crossing through chambem in H ER2-RNAi group [（63.6±5.1）/view3 was significantly lower than that in the control group[（193.5 ± 6.2）/view3 （P〈0. 05）. Conclusion HER2 silencing by RNAi suppresses the capacities of invasion and migration of human gastric cancer SGC-7901 cells, suggesting that RNAi gene silencing technique might be a new method in the treatment of advance gastric cancer.