RhoB对胃癌细胞SGC7901的负性调控作用

目的研究RhoB对胃癌细胞SGC7901增殖调控作用。方法用PCR扩增出RhoB的全长cDNA，构建RhoB可诱导表达载体pGene／V5-His-RhoB；脂质体介导法将调控载体pSwitch及诱导表达载体pGene／V5-His-RhoB先后转入人胃癌细胞SGC7901中，筛选出稳定抗性克隆；Western blot检测米非斯酮诱导后转染细胞中RhoB蛋白的表达。用MTT assay检测RhoB转染细胞株的生长速度、双苯并咪唑染料(Hoechst 33258)活细胞吸收分析法、流式细胞仪(FCM)检测RhoB对胃癌细胞凋亡指数作用。结果成功构建RhoB可诱导真核表达载体pGene／V5-His-RhoB，并经酶切和测序鉴定；转染后经潮霉素B和Zeocin筛选出稳定抗性克隆SGC7901／RhoB；米非斯酮诱导出RhoB蛋白表达，在诱导24小时后蛋白表达最高；细胞增殖试验结果显示，RhoB表达上调后胃癌细胞增殖受到抑制；细胞凋亡检测提示高表达RhoB可明显诱导胃癌细胞凋亡。结论RhoB对胃癌细胞增殖具有负性调控作用，可诱导胃癌细胞出现凋亡。

Suppressive effect of RhoB on gastric cancer cell line SGC7901

Objective To study the effect of RhoB on the proliferation of gastric cancer cell line SGC7901. Methods The inducible expression vector pGene/V5-His-RhoB containing the full length RhoB cDNA was constructed and transfected into gastric cancer cell SGC7901/pSwitch by lipofectamine, which was derived from SGC7901 after transfected with a regulatory plasmid pSwitch. The expression of RhoB in the screened stable cell lines were detected by Western blot after the induction of mifepristone for 0, 3, 6, 12, 24 hours. The effect of RhoB on the proliferation on gastric cancer cell line SGC7901 was evaluated by MTT assay. The apoptotic index was tested by Hoechst 33258 staining and FCM.Results The inducible expression vector pGene/V5-His-RhoB was constructed and confirmed by enzymatic digestion and DNA sequencing. After transfected into SGC7901 cells, RhoB protein expression, after mifepristone induction, was in a time dependence manner in 24 hours. MTT assay showed that RhoB suppressed growth of gastric cancer cells remarkably. Detection of apoptosis showed that RhoB enhanced apoptosis of the gastric cancer cells. Conclusion RhoB plays an important tumor-suppressive role in gastric cancer.