RNA干扰Nucleostemin基因对人结肠癌细胞株HT-29细胞增殖及细胞周期的影响  

Effect of Nucleostemin siRNA on cell proliferation and cell cycle in human colon cancer cell line HT-29

作　　者：王居峰[1] 刘莺[1] 刘文静[1] 何素英[1] 

机构地区：[1]河南省肿瘤医院肿瘤内科,郑州450003

出　　处：《华中师范大学学报（自然科学版）》2010年第3期463-467,共5页Journal of Central China Normal University：Natural Sciences

基　　金：中国临床肿瘤学科基金CSCO康莱特诺华基金(京ICP备05047300)

摘　　要：将Nucleostemin(NS)siRNA利用脂质体2000转染人结肠癌细胞株HT 29,分别利用CCK-8试剂盒和流式细胞术检测NS siRNA对HT 29细胞增殖和细胞周期的影响,进一步利用Real-time PCR和Western blotting技术检测NS基因和细胞周期相关基因p21 mRNA和蛋白的表达.结果表明,NS siRNA的转入能有效下调NS mRNA和蛋白的表达(P<0.05),并能明显抑制人结肠癌细胞株HT 29的细胞增殖(P<0.05),并诱导细胞周期静止在G_0/G_1期,转染NS siRNA后,p21 mRNA和蛋白的表达均明显升高,提示细胞增殖抑制和细胞周期静止与p21基因表达的升高密切相关.Nucleostemin （NS） siRNA was transfected into human colon cancer cell line HT-29 using LipofectamineTM 2000, and effects of NS siRNA on cell proliferation and cell cycle of HT-29 cells were detected by CCK-8 kit and flow cytometry, respectively. Further, the expressions of cell cycle related gene p21 mRNA and protein were investigated by Real-time PCR and Western blotting methods. The results demonstrated that introduction of NS siRNA into HT-29 cells effectively down-regulated the expression of NS mRNA and protein （P〈0.05）, and obviously inhibited cell proliferation of HT-29 cells （P〈0. 05）, and arrested cell cycle at G0/G1 phase, and the expressions of p21 mRNA and protein both elevated after transfection with NS siRNA, suggesting the inhibition of cell proliferation and cell cycle arrest may be closely associated with the elevation of the p21 gene.

关 键 词：核干细胞因子 结肠癌 细胞增殖 细胞周期 

分 类 号：Q78[生物学—分子生物学]