肿瘤细胞线粒体靶向表达Dm-dNK基因的实验研究  

Mitochondria targeted expression of drosophila melanogaster multisubstrate deoxyribonucleoside kinase in cancer cell lines

作　　者：赵蕾[1] 张鸿[2] 赵梅芬[3] 崔泽实[1] 李继光[4] 郑新宇[4] 

机构地区：[1]中国医科大学实验技术中心三部,辽宁沈阳1100012 [2]中国医科大学基础医学院,辽宁沈阳1100012 [3]中国医科大学第一附属医院普通外科,辽宁沈阳110001 [4]中国医科大学第一附属医院肿瘤外科,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2007年第9期641-644,共4页Chinese Journal of Cancer Prevention and Treatment

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

摘　　要：目的:探讨线粒体基质内核苷酸类似物的磷酸化水平,及Dm-dNK作为新的自杀基因杀伤肿瘤细胞的效果。方法:克隆线粒体信号于Dm-dNK序列N端,与绿色荧光蛋白(GFP)形成融合蛋白以确定基因的亚细胞水平表达位点,构建质粒转染骨肉瘤细胞及胰腺癌细胞,测定转染细胞Dm-dNK的活性和其对于嘌呤核苷酸类似物araC、dFdC、BVDU及araT的细胞毒性。结果:在所测试的细胞株中,线粒体基质内表达Dm-dNK具有较高的酶活性,转染肿瘤细胞对araC、araT及dFdC的敏感性增加。结论:Dm-dNK可以定位表达于肿瘤细胞线粒体并保持酶的活性,可能成为肿瘤基因治疗新的靶向作用位点。OBJECTIVE：To study the effects of nucleoside analog phosphorylation in the mitochondrial matrix, and killing effect using Dm-dNK as a novel suicide gene on cancer cell lines. METttODS： A Dm-dNK protein targeted to the mitochondrial matrix was created by fusing a mitochondrial targeting signal to the N-terminus of the protein, the constructed plasmids were transfected into osteosarcoma and pancreatic adenocarcinoma cells, and the enzymatic activity and the sensitivity of the untransfeeted cells and the cells transfeeted with either GFP vector alone or the mito Dm dNK-GFP to the pyrimidine nu cleoside analogs araC, dFdC, BVDU and araT were determined. RESULTS： The mitochondrial Dm-dNK was enzymatically active and the overexpression of the enzyme in the tested cell lines resulted in an increased sensitivity to the cytosine nucleoside analogs such as araC, ar aT and dFdC. CONCLUSIONS： Dm dNK could be locally overcx pressed to mitochondria and it retaines enzymatic activity. It may con tribute to the development of novel 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 DNA 线粒体 

分 类 号：R73-3[医药卫生—肿瘤]