TNF-α基因转染多药耐药性人乳腺癌细胞的基因表达与生物学效应  

Expression and Biological Effects of TNF-α Gene Transfected into Multidrug Resistant Human Breast Cancer Cell Line

作　　者：郭伟剑[1] 沈兆忠[2] 钱关祥[3] 罗建明[2] 郑颂国[2] 周振华[2] 

机构地区：[1]上海第二医科大学新华医院肿瘤中心,200092 [2]上海医科大学肿瘤医院,200032 [3]上海第二医科大学人类基因治疗研究中心,200025

出　　处：《实用癌症杂志》2000年第6期575-577,共3页The Practical Journal of Cancer

摘　　要：目的　探索TNF α基因能否成功导入耐药细胞并获得表达 ,发挥生物学效应。方法　以重组逆转录病毒为载体将TNF α基因导入具有多药耐药 (MDR )表型的人乳腺癌细胞系MCF7/ADR ,经G 418抗性筛选获 4株阳性克隆。PCR、Elisa法鉴定并检测TNF α基因的整合和表达。细胞计数法观察细胞生长速度的改变 ,流式细胞仪检测细胞周期变化。结果　 4株阳性克隆整合了TNF α基因并获得表达 ,分泌TNF α量分别为 341pg/ml(10 6cell/48h)、5 40 pg/ml、1737pg/ml、2 875 pg/ml。与对照细胞相比 ,TNF α分泌量较低的 2株细胞克隆生长速度无明显减慢 ,而 2株高表达TNF α的细胞生长速度明显减慢 ,生长抑制率分别为 32 .4%、5 4.8% ,且细胞周期受阻于S +G2 /M期。结论　TNF α基因能成功导入耐药细胞并获得表达。高表达TNF α的细胞克隆生长明显受抑。Objective To study if TNF-α gene can be transfected into multidrug resistant cell line and has biological effects.Methods Via a recombinant retrovirus vector,TNF-α gene was transfected into multidrug resistant human breast cancer cell line MCF7/ADR.After G418 screening,4 positive clones were obtained.The integration and expression of TNF-α gene were analyzed by PCR and ELISA.The biological effects of TNF-α were investigated by cells account and flow cytometric assay.Results The integration and expression of TNF-α gene were successful.The secreting level of TNF-α in the 4 TNF-α gene trasfected cell clones were 341 pg/ml(106 cell/48 h),540 pg/ml,1 737 pg/ml,and 2 875 pg/ml,respectively.In comparison with control,the 2 latter cell clones which secreting high level of TNF-α showed lower growth rate(the growth inhibiting rate were 32.4%,and 54.8%,respectively),and its′ cell cycle was arrested at S and G 2/M phase.Conclusion TNF-α gene can be transfected into drug resistant cell.The growth of cell clone expressing high level TNF-α is inhibited.

关 键 词：乳腺癌 肿瘤坏死因子 基因治疗 多药耐药 生物学 

分 类 号：R737.905[医药卫生—肿瘤] R730.59[医药卫生—临床医学]