Tie2-siRNA表达载体对子宫内膜癌Ishikawa细胞株化疗敏感性的研究  

Reduced Tie2 expression increases the chemosensitivity of the Ishikawa endometrial cancer cell line to Carboplatin

作　　者：吴婵[1] 周怀君[1] 寻庆英[2] 丁利军[1] 

机构地区：[1]南京大学附属鼓楼医院,南京210000 [2]东南大学临床医学院,南京210000

出　　处：《现代妇产科进展》2015年第12期885-888,共4页Progress in Obstetrics and Gynecology

摘　　要：目的:研究Tie2表达对子宫内膜癌细胞株化疗敏感性的影响,探索子宫内膜癌辅助治疗新方法。方法:将表达Tie2-siRNA的碱基片段插入含有CMV启动子、新霉素抗性基因及绿色荧光蛋白报告基因的质粒中,应用脂质体包涵体技术将质粒转染入子宫内膜癌Ishikawa细胞株,以减少Tie2基因表达,采用逆转录PCR及蛋白印迹方法鉴定。DAPI染色检测各组细胞凋亡。不同浓度卡铂处理细胞,检测各组细胞的化疗敏感性。结果:成功转染后,Ishikawa细胞中Tie2 mRNA及蛋白表达水平的抑制率分别为75.5%及56.4%,Tie2-siRNA组细胞凋亡增加,化疗敏感性增加。Tie2-siRNA组的卡铂IC50为(29.95±0.68)μg/ml,低于空白对照组(80.55±1.05)μg/ml。结论:降调Tie2基因的转录及表达可促进子宫内膜癌细胞凋亡,增加子宫内膜癌细胞对化疗药物的敏感性。Objective：In this study, we tested the effect of Tie2 gene expression on the chemosensifivity of the Ishikawa human cndometrial cancer cell line. The aim of this study is to explore the new auxiliary treatment method for endometrial cancer. Methods：The Tie2-siRNA sequence was cloned into a plasmid containing the CMV promoter, the neomycin resistance gene, and the green fluorescent protein （GFP） gene. To reduce Tie2 expression, the Ishikawa endometrial cancer cell line was transfeeted with the recombinant plasmid using Lipofectamine 2000. Then, reverse transcription PCR （RT-PCR） and western blotting was used to analyze Tie2 mRNA and protein levels. We also assessed the effects of Tie2-silcneing on morphological changes in the Ishikawa cells. The chemosensitivity of transfected Ishikawa cells was investigated using carboplatin-induced cell death and methyl thiazolyl tetrazolium （MTT） detection. Resuits：The Tie2 mRNA transcript and protein expression were reduced in the transfected Ishika- wa cells by 75.5% and 56.4% ,respectively. The data show there was increased cell apoptosis and enhanced chemosensitivity in the Tie2-siRNA Ishikawa cells. The IC50 of carboplatin was （29.95 ±0.68 ）μg/ml in the Tie2-siRNA group and （80.55± 1.05 ）pog/ml in the control group. Conclusion： The results of this study indicate that cell apoptosis and ehemosensitivity can be increased by inhibiting Tie2 expression in the Ishikawa cell line.

关 键 词：TIE2 子宫内膜癌 Ishikawa细胞株 小干扰RNA 

分 类 号：R737.33[医药卫生—肿瘤]