稳定表达GFP-C-Daam1的MDA-MB-231人乳腺癌细胞系的建立  

Establishment of a stable GFP-C-Daam1 MDA-MB-231 breast cancer cell line

作　　者：李卫星[1] 杜军[1] 胡圳圳[1] 田寅辉[1] 刘皎婧[1] 朱一超[1] 

机构地区：[1]泰州职业技术学院医学技术学院,江苏泰州225300

出　　处：《南京医科大学学报（自然科学版）》2011年第7期925-929,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(30872926);江苏省博士后科研资助(0902023C);南京医科大学科技发展基金重点项目(2010NJMUZ35)

摘　　要：目的:构建稳定表达GFP和GFP-C-Daam1的MDA-MB-231人乳腺癌细胞系,以便进一步研究Daam1对乳腺癌细胞迁移能力的影响。方法:构建表达GFP和GFP-C-Daam1的慢病毒载体,通过慢病毒感染获取稳定表达目的基因的MDA-MB-231人乳腺癌细胞系。用嘌呤霉素筛选稳定表达的细胞系,Western blot确定重组蛋白表达,并用Boyden chamber小室实验检测细胞运动能力的改变。结果:通过慢病毒感染,建立了稳定表达GFP-C-Daam1的MDA-MB-231人乳腺癌细胞系,该细胞系具备更强的内在运动能力,表明目的基因的表达产物功能正常。结论:用慢病毒载体可以构建稳定表达GFP-C-Daam1的MDA-MB-231人乳腺癌细胞系,从而得到研究活化的Daam1的可靠细胞膜型。Objective：To establish a MDA-MB-231 human breast cancer cell line stably express GFP-C-Daam1,and to investigate the influence of Daam1 on the mobility of breast cancer cells.Methods：The lentivrial vectors containing GFP or GFP-C-Daam1 were constructed and infected human breast cancer cells MDA-MB-231.MDA-MB-231 cells were screened by puromycin and stable expression of GFP or GFP-C-Daam1 was confirmed by Western blotting.Boyden chamber assay was used to test the intrinsic motility of the cells.Results：The cell lines stably expressing GFP or GFP-C-Daam1 were established successfully by the lentivrial infection.The GFP-C-Daam1 expressing cells showed greater intrinsic motility,suggesting the products of the interesting gene were active and functional.Conclusion：MDA-MB-231 breast cancer cell lines stably expressing GFP or GFP-C-Daam1 were successfully established using lentiviral methods.These cell lines can be used as model cells for further study of active Daam1 targeting.

关 键 词：C-Daam1 MDA-MB-231细胞 基因转染 稳定表达 细胞迁移 

分 类 号：R730.23[医药卫生—肿瘤]