摘要
利用RNA干扰技术沉默结肠癌细胞株HT-29细胞中Nucleostemin(NS)基因的表达,并分析其对HT-29细胞凋亡的影响,进一步利用半定量RT-PCR和Western blotting技术检测细胞凋亡相关基因caspase-3/9 mRNAs和蛋白的表达.结果表明,NS基因的沉默能明显提高caspase-3/9的活性(P<0.05),并诱导细胞发生凋亡,此外,HT-29细胞中NS基因沉默后,caspase-3/9mRNAs和蛋白的表达均明显升高,提示NS基因沉默介导的细胞凋亡与caspase-3/9基因表达的升高密切相关.
Nucleostemin (NS) gene expression was silenced in human colon cancer cell line HT-29 using RNA interference technology, and effects of knock-down of NS gene on cell apoptosis of HT-29 cells were analyzed by caspase activity and flow cytometry. Further,the expressions of cell apoptosis related gene caspase-3/9 mRNAs and proteins were investigated by Real-time PCR and Western blotting methods. The results demonstrated that knock-down of NS gene evidently elevate caspase-3/9 activities (P〈0.05), and induced cell apoptosis of HT-29 cells,in addition,expressions of caspase-3/9 mR- NAs and proteins significantly improved after transfection with NS siRNA, suggesting cell apoptosis mediated by knock-down of NS gene may be closely associated with the elevation of the caspase-3/9 genes.
出处
《华中师范大学学报(自然科学版)》
CAS
CSCD
北大核心
2010年第1期125-128,共4页
Journal of Central China Normal University:Natural Sciences
基金
中国临床肿瘤学-康莱特-诺华基金(京ICP备05047300)