逆转录病毒介导Dm-dNK基因在乳腺癌细胞中的表达及细胞杀伤毒性  

Retrovirus-mediated expression of Dm-dNK in breast cancer cells and its cytotoxicity

作　　者：赵蕾[1] 郑新宇[2] 张鸿[3] 李继光[2] 徐惠绵[2] 

机构地区：[1]中国医科大学实验技术中心三部,沈阳110001 [2]中国医科大学第一附属医院乳腺外科,沈阳110001 [3]中国医科大学基础医学院,沈阳110001

出　　处：《中华乳腺病杂志（电子版）》2007年第5期165-167,共3页Chinese Journal of Breast Disease(Electronic Edition)

基　　金：国家自然科学基金(30371625);"863"计划(2006AA02Z493)资助项目

摘　　要：目的探讨可否将果蝇多底物脱氧核苷酸激酶(Dm-dNK)表达于乳腺癌细胞,以及其对肿瘤细胞的杀伤效果。方法构建表达Dm-dNK复制缺陷的逆转录病毒载体;重组病毒感染乳腺癌细胞系MCF7(ER+)及MDA-MB-231(ER-),测定感染细胞酶的活性及对于核苷酸类似物araT及araC的细胞毒性。结果 Dm-dNK可以在乳腺肿瘤细胞中表达并具有酶的活性(Western印迹显示细胞内的Dm-dNK蛋白表达);同时,感染的肿瘤细胞对araT及araC的敏感性增加。结论 Dm-dNK可以定位表达于乳腺肿瘤细胞核并保持酶的活性,Dm-dNK可能成为乳腺肿瘤分子化疗的新方法。Objective To evaluate the effects of nucleoside analogphosphorylation by Dm-dNK （Drosophila melanogaster deoxyribonucleosidekinase）, and the killing effect as a novel suicide geneon in breast cancer cell lines. Methods A replication-deficientretroviral vector that expresses Dm-dNK was created, and the human breast cancer cells MCF7 （ER ＋ ） and MDA-MB-231 （ER - ） were transduced with the recombinant retrovirus. The enzymatic activity and the sensitivity of the untransfected cells and the cells infected with either GFP （green fluorescent protein） vector alone or the pLE-Dm-dNK-GFP to pyrimidine nucleoside analogs araT （1-β-Darabinofuranosyhhymine） and araC （1-β-D-arabinofuranosylcytosine） were determined. Results It was showed that the Dm-dNK was enzymatically active and that the over-expression of the enzyme in the tested cell lines resulted in an increased sensitivity to the cytosine nucleoside analogs araT and araC. Conclusions Dm-dNK can be locally overexpressed in the nuclei of breast cancer cells and it retains enzymatic activity. It may contribute to the development of novel breast cancer treatment strategies.

关 键 词：基因治疗 分子化疗 核苷酸激酶 核苷酸类似物 

分 类 号：R737.9[医药卫生—肿瘤]