慢病毒介导Survivin shRNA与结肠腺瘤息肉易感基因片段联合对HT-29细胞的影响  

Effect of lentiviral-mediated delivery of Survivin sh RNA with adenomatous polyposis coli fragment in HT-29 cells

作　　者：袁禧先[1] 隋子奇 孙理婷 杨延涛[2] 张爽[3] 薛鸿鹏[4] 葛文松[5] 王凤荣[6] 

机构地区：[1]佳木斯大学附属第一医院消化二科,黑龙江省佳木斯市154003 [2]佳木斯大学临床医学院,黑龙江省佳木斯市154003 [3]齐齐哈尔市第一医院消化科,黑龙江省齐齐哈尔市161005 [4]黑龙江省医院消化内科,黑龙江省哈尔滨市150030 [5]上海交通大学医学院附属新华医院消化科,上海市200000 [6]佳木斯大学附属第一医院研究生办公室,黑龙江省佳木斯市154003

出　　处：《世界华人消化杂志》2015年第14期2250-2255,共6页World Chinese Journal of Digestology

基　　金：2014年佳木斯大学研究生科技创新基金资助项目;No.LM2014_030;黑龙江省2011年研究生创新科研资金资助项目;No.YJSCX2011-380HLJ;黑龙江省自然科学基金资助项目;No.H201368~~

摘　　要：目的:观察Survivin sh RNA、结肠腺瘤息肉易感基因(adenomatous polyposis coli,APC)片段联合对HT-29细胞Survivin表达及细胞增殖的影响.方法:构建Survivin sh RNA慢病毒载体、A P C有效片段慢病毒载体,对H T-29细胞分别采用单慢病毒载体侵染及联合侵染.实验分阴性对照组、空载组、sh RNA组、APC组、sh RNA+APC联合组,对侵染48 h后的HT-29细胞进行real-time PCR、Western blot及CCK8细胞增殖检测,检测Survivin m RNA、蛋白表达水平及对细胞增殖的影响.结果:(1)sh RNA+APC联合组与其余各组相比,Survivin m RNA表达量显著下降(P<0.05);(2)s h R N A+A P C联合组与其余各组相比,其Survivin蛋白抑制率明显高于其余各组(P<0.05);(3)sh RNA+APC联合组与其余各组相比,细胞增殖速度低于其余各组(P<0.05).结论:Survivin sh RNA与APC片段联合能抑制HT-29细胞内Survivin m RNA及蛋白的表达,同时能够抑制细胞增殖能力,并且优于单个基因侵染.AIM： To observe the effect of Survivin sh RNA combined with adenomatous polyposis coli（APC） fragment on the expression of Survivin and cell proliferation in HT-29 cells.METHODS： Lentiviral vectors for Survivin sh RNA and APC fragment were constructed and transfected into HT-29 cells, alone or in combination. Cells were divided into a negative control group, an empty vector group, an shR NA group, an APC group, and an shR NA ＋ APC group. After 48 h of infection, the Survivin m RNA and protein expression was detected by real-time PCR and Western blot, respectively. The proliferation of cells was assayed by CCK8.RESULTS： ShR NA ＋ APC significantly decreased the expression of Survivin m RNA and protein and cell proliferation compared with the other groups（P〈0.05 for all）. CONCLUSION： Survivin sh RNA combined with APC fragment can inhibit the expression of Survivin m RNA and protein and downregulate cell proliferation in HT-29 cells.

关 键 词：结肠癌 SURVIVIN SHRNA APC有效片段慢病毒载体 

分 类 号：R735.35[医药卫生—肿瘤]