Tie2-siRNA表达载体对子宫内膜癌Ishikawa细胞株生物学行为的影响  

Effects of Tie2-siRNA expression vector on biological behavior of human endometrial Ishikawa cell line

作　　者：吴婵[1] 周怀君[1] 寻庆英[2] 李荣[1] 

机构地区：[1]南京大学医学院附属鼓楼医院妇产科,江苏省210008 [2]东南大学基础医学院

出　　处：《江苏医药》2016年第8期879-882,F0002,共5页Jiangsu Medical Journal

基　　金：南京市医学科技发展项目(ZKX09023)

摘　　要：目的探讨Tie2-siRNA表达载体对子宫内膜癌Ishikawa细胞株生物学行为的影响。方法将表达Tie2-siRNA的碱基片段插入含有巨细胞病毒启动子、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基因的表达,并采用逆转录PCR及Western blot方法鉴定。用MTT实验描绘各组细胞增殖曲线,流式细胞术检测各组细胞凋亡率。结果成功转染后Tie2基因的mRNA转录水平及蛋白表达水平抑制率分别为75.5%及56.4%,Ishikawa细胞增殖72-h抑制率为73.9%,凋亡率为35.9%。结论体外下调Tie2基因的转录及表达可抑制肿瘤生长,促进细胞凋亡。Objective To investigate the effects of short interfering RNA（siRNA）targeting Tie2（Tie2-siRNA）expression vector on biological behavior of human endometrial Ishikawa cell line.Methods A plasmid based on cytomegalovirus promoter,neomycin resistance gene and green fluorescent protein gene was used to deliver and express siRNA targeting Tie2 to reduce its expression in Ishikawa cells.RT-PCR and Western blot analysis were used to measure the mRNA and protein levels of Tie2.The effects of Tie2-silencing on Ishikawa cells were analyzed by a cell growth curve and apoptosis test.Results The expression level of Tie2 mRNA in Ishikawa cells transfected with siRNA was decreased by 75.5%,and protein level was decreased by 56.4%.After transfecting,the inhibitory rate of proliferation after 72 hours and apoptosis rate in Ishikawa cells were 73.9% and 35.9%,respectively.Conclusion The downregulation of Tie2 in vitro results in significant inhibition of tumor growth and promotion of the apoptosis.

关 键 词：Tie2基因 子宫内膜癌 Ishikawa细胞株 小干扰RNA 

分 类 号：R737[医药卫生—肿瘤]