Livin基因和Survivin基因联合靶向siRNA抑制大肠癌细胞增殖的作用  

Inhibitory effect of multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes on human colon cancer cells proliferation

作　　者：蔡明[1] 王国斌[1] 陶凯雄[1] 蔡昌学[2] 

机构地区：[1]华中科技大学同济医学院附属协和医院普通外科,湖北武汉430022 [2]华中科技大学同济医学院基础医学院病原生物系,湖北武汉430030

出　　处：《药学学报》2009年第7期741-746,共6页Acta Pharmaceutica Sinica

基　　金：国家高技术研究发展计划(863计划)资助项目(2001AA218051);湖北省科技攻关计划资助项目(2005A304B09)

摘　　要：采用1个载体编码2个shRNA技术,构建Livin基因和Survivin基因联合靶向的siRNA重组表达载体,研究其对人大肠癌细胞生长协同抑制的增效作用。构建Livin基因和Survivin基因联合靶向的siRNA重组表达载体并转染大肠癌细胞,通过RT-PCR和Western blotting方法检测Livin基因与Survivin基因的表达,利用流式细胞仪检测处理后细胞的凋亡效应。经酶切鉴定和测序分析证实Livin基因和Survivin基因联合靶向的siRNA重组表达载体构建成功,它对大肠癌细胞Livin和Survivin mRNA的抑制率分别为27.90%和32.24%,对Livin和Survivin蛋白表达的抑制率分别为22.28%和40.86%,诱导肿瘤细胞凋亡率为(11.69±1.37)%,但协同干扰作用比单独干扰Livin基因或Survivin基因弱。Livin基因和Survivin基因联合靶向的siRNA重组表达载体构建成功并能抑制Livin基因和Survivin基因的表达,但协同抑制作用比单独干扰Livin基因或Survivin基因弱。In order to maximize the efficiency and versatility of the vector-based siRNA approach, we have developed a novel siRNA expression vector containing multiple tandem siRNA cassettes to investigate the synergistic inhibitory effect of it on human colon cancer cells proliferation. Multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed and transfected into human colon cancer cell. The effect of multiple siRNA recombinant expression vector was detected by RT-PCR, Western blotting and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed successfully. Livin and Survivin genes inhibition ratio of Livin and Survivin siRNA at mRNA levels were 27.90% and 32.24%, at protein levels were 22.28% and 40.86%, the apoptotic ratio was （11.69 ± 1.37） %, but the synergistic effect was weaker than Livin and Survivin RNA interference, respectively. The multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes has been constructed successfully. It can inhibit the expression of Livin and Survivin genes in human colon cancer ceils, but the synergistic effect was weaker than Livin and Survivin RNA interferences, respectively.

关 键 词：LIVIN基因 SURVIVIN基因 RNA干扰 基因治疗 

分 类 号：R963[医药卫生—微生物与生化药学]