稳定表达siRNA-ARK5基因的胃癌SGC-7901细胞系的构建  

Establish gastric cancer SGC-7901 cell line with stable siRNA-ARK5 gene expression

作　　者：李文花 王海超[2] 雷丽恰[2] 张月超[2] 史立宏[2] 张宝刚[3] 

机构地区：[1]潍坊市第二人民医院消化内科,山东潍坊261042 [2]潍坊医学院药理学实验室 [3]潍坊医学院病理学教研室

出　　处：《青岛医药卫生》2013年第5期321-323,329,共4页Qingdao Medical Journal

基　　金：国家自然科学基金资助项目(81072068);山东省自然科学基金(ZR2011HL047);潍坊市科学技术发展计划项目(201201230)

摘　　要：目的构建稳定表达siRNA-ARK5基因的人胃癌SGC-7901细胞系,为研究ARK5在胃癌发生发展中的作用与机制奠定基础。方法把pGCsilencerU6/GFP/Neo-RNAi-ARK5重组质粒通过脂质体转染胃癌SGC-7901细胞,利用不同浓度的G418筛选获得稳定转染细胞株,挑取单克隆细胞系进行培养,应用荧光显微镜和RT-PCR技术确定是否获得了ARK5基因被稳定抑制的单克隆细胞系。结果成功挑取的4株单克隆细胞中2株细胞的ARK5基因的表达被显著抑制。结论通过siRNA干扰技术成功构建了稳定表达siRNA-ARK5基因的胃癌SGC-7901细胞系,为研究ARK5在胃癌发生发展中的作用与机制奠定了基础。Objective To establish gastric cancer SGC-7901 cell lines that stably express siRNA-ARK5 for the study of the mechanism of ARK5 on the occurrence and development of gastric cancer. Methods The pGCsilencerU6/GFP/Neo-RNAi-ARK5 plasmids were transfected into gastric cancer SGC-7901 cells with lipofectamine. G418 was used to selected out stable transfection clone cells, the efficiency of transfection and ARK5 gene silencing were evaluated by RT- PCR and fluorescence microscopy. Results ARK5 gene expression was significantly inhibited in 2 of 4 selected stable clones. Conclusion We successfully established gastric cancer SGC-7901 cell line with stable siRNA-ARK5 gene expression. This cell line can provide a cell model for investigating the relationship between ARK5 and the gastric cancer malignancy.

关 键 词：胃癌 ARK5 SIRNA 

分 类 号：R730.2[医药卫生—肿瘤]