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目的构建稳定转染人组织因子(TF)的人胃癌细胞株SGC7901,研究TF对胃癌细胞体外侵袭转移特性的影响。方法采用巢式PCR技术自人胎盘组织克隆目的基因TF,应用脂质体介导的转染技术将其导入人胃癌细胞株SGC7901,采用G418筛选稳定表达TF的细胞,采用RT-PCR及Western blot法检测TF mRNA及蛋白的表达,并进行Transwell实验及划痕实验观察细胞体外侵袭转移能力的变化。结果经基因测序证实成功构建了稳定、高效表达TF的人胃癌细胞系SGC7901/TF。转染组TF mRNA及TF蛋白表达增高;与阴性对照组及空白对照组相比,转染组细胞的体外侵袭转移能力增强。结论 TF能够增强人胃癌细胞的体外侵袭转移能力。
Objective To construct a human gastric cancer cell line SGC7901 stably transfected with human tissue factor (TF) to study the effect of TF on invasion and metastasis of gastric cancer cells in vitro. Methods The gene TF was cloned from human placenta by nested PCR and introduced into human gastric cancer cell line SGC7901 by liposome-mediated transfection. The cells stably expressing TF were screened by G418. The expression of TF was detected by RT-PCR and Western blot Method to detect the expression of TF mRNA and protein. Transwell experiments and scratching experiments were performed to observe the changes of invasion and metastasis of cells in vitro. Results The human gastric cancer cell line SGC7901 / TF stably and efficiently expressing TF was successfully constructed by gene sequencing. Transfection group TF mRNA and TF protein expression increased; compared with the negative control group and blank control group, transfected cells in vitro invasion and metastasis increased. Conclusion TF can enhance the invasion and metastasis of human gastric cancer cells in vitro.