RNAi沉默CD133基因表达对结肠癌细胞株HT-29影响的研究  

Inhibition of CD133 by RNA Interference in human colon carcinoma cell line HT-29

作　　者：唐茂盛[1] 王伟军[1] 彭欢[1] 姚厚山[1] 胡志前[1] 

机构地区：[1]第二军医大学附属长征医院普外科,上海200003

出　　处：《中华肿瘤防治杂志》2011年第12期936-939,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：上海科委科研计划项目(08140902900)

摘　　要：目的:探讨通过RNA干扰沉默CD133基因对结肠癌细胞株HT-29的增殖和侵袭力的影响。方法:将CD133siRNA用脂质体转染入HT-29细胞,以逆转录-聚合酶链反应(RT-PCR)检测CD133mRNA的变化,蛋白质印迹法检测转染前后蛋白的表达水平,用MTT比色法和Transwell小孔实验分别检测癌细胞的增殖及侵袭力变化。结果:CD133siRNA转染成功,与正常对照组相比,转染组CD133mRNA及蛋白的表达水平均有明显的抑制,并呈现浓度时间依赖性。MTT实验证明转染组细胞随时间延长,增殖逐渐受到抑制,P<0.05。Tran-swell小孔实验中经转染处理的穿膜细胞数明显减少(P<0.05),RNA干扰降低了细胞的侵袭性。结论:CD133在HT-29癌细胞增殖及侵袭过程中起重要作用,RNA干扰可以有效抑制其CD133的表达,并能有效降低癌细胞的增殖及侵袭力。OBJECTIVE： To construct an expression plasmid of siRNA against CD133 gene and to investigate its biological behavioral effect on human colon carcinoma cell line HT-29. METHODS： we inhibited the CD133 expression using the siRNA method. Expression levels of CD133 mRNA and protein from HT 29 after transfection were examined by RT-PCR and Western blot respec tively. Changes of the cell growth activity in response to transfect ed plasmid were evaluated by MTT assay. The migration ability of transfected HT-29 cells were measured quantitatively by Transwell migration assays. RESULTS： After CD133 siRNA transfeetion into HT-29 cells, compared with that in control groups,the level mRNA of CD133 in cancer cells transfected with CD133 siRNA was inhibited in a concentration and time dependent manner and the CD133 protein con- centration decreased significandy. The growth of HT29 cells was sig nificantly inhibited than those in control groups （P〈 0.05）. CD133 siRNA significandy inhibited invasion of HT29 cells to matrigel （P〈0. 05）. CONCLUSION： Silencing CD133 gene by the siRNA technology can actively suppress the expression of CD133 gene, and then inhibit the growth and invasion of HT-29 cell.

关 键 词：大肠肿瘤 RNA干扰 基因 CDl 33 基因治疗 HT-29细胞 

分 类 号：R735.35[医药卫生—肿瘤]