外源性sall4基因在胃癌HGC27细胞株中的表达及鉴定

Enforced expression and verification of sall4 in HGC27 cell line

目的:建立sall4稳定转染的HGC27细胞株,为sall4基因在胃癌发生发展中的作用研究奠定基础。方法:采用脂质体介导法将pCMV6-entry-sall4及pCMV6-entry质粒转染至HGC27细胞,48 h后加入终质量浓度600mg/L的G418抗生素进行药物加压筛选,获得阳性单克隆细胞株,通过RT-PCR和蛋白质印迹检测细胞中sall4的表达情况。结果:成功构建了稳定过表达sall4的HGC27细胞株,稳定转染细胞株中sall4的mRNA和蛋白水平明显高于未转染组及转染空载体组。结论:本实验成功筛选出sall4基因过表达的稳定转染胃癌细胞系。

Objective： To establish the stable sall4-overexpressing HGC27 cell line,so as to lay a foundation for exploring the role of sall4 in the development of gastric cancer.Methods： pCMV6-entry-sall4 and pCMV6-entry plasmids were transfected into HGC27 cells via Lipofectamine 2000.At 48 h after transfection,cells were then cultured with G418 at a final concentration of 600 mg / L.Finally,stably transfected sublines of HGC27 cells were obtained.The expression of sall4 in stable cell line was determined by RTPCR and western blotting.Results： The HGC27-sall4 cell line was obtained.The expression level of SALL4 in HGC27-sall4 cells was significantly higher than untransfected HGC27 cells and HGC27-vector cells.Conclusion： We successfully screened out the stable sall4-overexpressing HGC27 cell line.