论文部分内容阅读
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 observe the effect of exogenous RASSF1A gene on the proliferation of human gastric cancer cell line SGC7901. Methods Eukaryotic expression recombinant pcDNA3.0-RASSF1A plasmid and empty vector pcDNA3.0 plasmid were transfected into SGC7901 cells by lipofectamine transfection, and stable transfected cell clones were obtained after selection by G418. The expression of RASSF1A gene was detected by RT-PCR and Western blot. The cell growth curve was plotted, and the biological behavior of the transfected cells was analyzed using a nude mouse tumor formation assay, a plate colony formation assay, and flow cytometry. Results The SGC7901 cell line with high expression of RASSF1A gene was established by liposome transfection and screening. Compared with the untransfected group and the transfected blank vector group, the growth rate of SGC7901 cells transfected with RASSF1A gene was significantly slower; the ratio of G1/G0 phase in the cell cycle was significantly increased, while the proportion in S phase was decreased; the cloning rate was significantly reduced; Rat tumor inhibition rate was 57.1%. Conclusion RASSF1A gene can inhibit the proliferation of human gastric cancer cell line SGC7901.