文章摘要
李卫星,杜 军,胡圳圳,田寅辉,刘皎婧,朱一超.稳定表达GFP-C-Daam1的MDA-MB-231人乳腺癌细胞系的建立[J].南京医科大学学报,2011,(7):925~929
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
投稿时间:2011-02-19  
DOI:10.7655
中文关键词: C-Daam1  MDA-MB-231细胞  基因转染  稳定表达  细胞迁移
英文关键词: C-Daam1  MDA-MB-231 cell  gene transfection  stable expression  cell migration
基金项目: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)
作者单位
李卫星 泰州职业技术学院医学技术学院,江苏 泰州 225300 
杜 军 南京医科大学生理学系,江苏 南京 210029 
胡圳圳 南京医科大学生理学系,江苏 南京 210029 
田寅辉 南京医科大学生理学系,江苏 南京 210029 
刘皎婧 南京医科大学生理学系,江苏 南京 210029 
朱一超 南京医科大学生理学系,江苏 南京 210029 
摘要点击次数: 1550
全文下载次数: 146
中文摘要:
      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?
英文摘要:
      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.
查看全文   查看/发表评论  下载PDF阅读器