RNA干扰技术对大肠癌细胞人端粒酶逆转录酶基因的抑制作用  

Inhibitory effects of RNA interference on hTERT gene in colon cancer HT-29 cells

作　　者：袁超君[1] 李其云[1] 易波[1] 黎才海[1] 吴亮高[1] 

机构地区：[1]江西省肿瘤医院腹部外科,南昌330029

出　　处：《中华实验外科杂志》2009年第4期464-466,共3页Chinese Journal of Experimental Surgery

基　　金：江西省卫生厅学科带头人基金资助项目（20045050）

摘　　要：目的 观察RNA干扰对大肠癌细胞中人端粒酶逆转录酶基因（hTERTgene）的抑制效应。方法大肠癌细胞株HT-29分为Control组、N—hTERT组、Lipofectamine组和sihTERT组；将靶向于hTERT基因的小干扰RNA，转染大肠癌细胞；分别检测大肠癌细胞转染前后的端粒酶活性，hTERTmRNA和蛋白水平，细胞生长增殖和凋亡情况。结果sihTERT转染大肠癌细胞效率为60％；sihTERT组的端粒酶活性值、hTERTmRNA表达、hTERT蛋白表达、细胞凋亡率分别为0．73±0．14、0．47±0．08、0．37±0．07、49．5％，sihTERT组与其他三组比较差异有统计学意义（P〈0．01）。结论靶向于hTERT的siRNA能特异性沉默大肠癌细胞hTERT基因，下调hTERT基因mRNA及蛋白表达水平，降低端粒酶活性，抑制癌细胞生长增殖从而促进大肠癌细胞凋亡。Objective To explore the inhibitory effect of RNA interference on the human telomerase reverse transcriptase gene in colon cancer cells. Methods Colon cancer HT-29 cells were divided into control group,N-hTERT group, Lipofectamine group, and sihTERT group. The activity of telomerase, and expression of hTERT mRNA and protein were detected before and after siRNA was transfected into HT-29 cells. Flow cytometry was used to observe the proliferation and apoptosis of HT-29 cells. Results The transfection efficiency of siRNA was 60%. The activity of telomerase, the expression levels of hTERT mRNA and protein,and apoptosis rate in sihTERT group were 0.73 ± 0. 14,0.47 ± 0.08,0.37 ±0.07, and 49.5% respectively, which were significantly different from those in other three groups （P 〈 0.01 ）. Condusion siRNA targeting hTERT can speeificly scilence the hTERT gene of colon cancer cells by downregulating the expression of hTERT mRNA and protein, and suppressing the telomerase activity. RNAi can inhibit the proliferation of the cancer cells and induce their apoptosis.

关 键 词：大肠癌 端粒酶逆转录酶 RNA干扰 

分 类 号：R735.34[医药卫生—肿瘤] R73[医药卫生—临床医学]