RASSF1A对人胃癌细胞株SGC7901增殖的影响

The effect of RASSF1A gene on the proliferation of human gastric carcinoma cell line SGC7901

目的观察外源性RASSF1A基因对人胃癌细胞SGC7901增殖的影响。方法利用脂质体转染技术将真核表达重组体pcDNA3.0-RASSF1A质粒和空载体pcDNA3.0质粒分别导入SGC7901细胞,经G418筛选后获得稳定转染细胞克隆。采用RT-PCR和蛋白印迹检测RASSF1A基因表达。绘制细胞生长曲线,并用裸鼠成瘤实验、平板克隆形成实验、流式细胞术分析转染细胞的生物学行为。结果经脂质体转染和筛选,建立了高表达RASSF1A基因的SGC7901细胞系。与未转染组和转染空白载体组比较,转染RASSF1A基因的SGC7901细胞生长速度明显减慢;细胞周期中G1/G0期比例明显增加,而S期比例减少;克隆形成率明显减少;裸鼠成瘤抑制率为57.1%。结论RASSF1A基因能抑制人胃癌细胞SGC7901的增殖。

Objective To study the effects of exogenous RASSF1 A gene on the proliferation of gastric carcinoma cell line SGC7901. Methods The mammal expression vector pcDNA3. 0-RASSFIA and pcDNA3.0 were introduced into SGC7901 cell line by lipofectin transfection, and the SGC7901 cells stably overexpressing RASSF1 A gene were established by G418 selection. The expression of RASSF1 A gene was detected by Western blotting and RT-PCR. The cytobiologic characterizations of the positive clone were analyzed by MrlT assay, eytometry, colony assay, and tumorigenicity experiment. Results SGC7901 cells with stably expression RASSF1 A protein were established by lipofection mediated transfection and selected for further study. Compared with the non-transfected and vector transfected ceils, the positive clone cells developed more slowly; flow eytometric data showed that more positive clone cells went into phase G0/G1 and less cells went into phase S （P〈0.05） ; colony formation efficiency of the positive clone cells was lower than that of non-transfected and vector transfected cells（P〈0.05）. The average weight of tumor tissue in nude mice in positive clone cells was less than that of non-transfected and vector transfected cells （P〈0.05）. Conclusions RASSF 1 A gene can suppress the proliferation of SGC7901 cell line.