survivin靶向SiRNA协同5-FU对结直肠癌细胞的化疗增敏作用  

The effect of enhancement chemotherapy sensitivity by combining SiRNA recombinant expression vector targeting survivin gene with 5-Fu on human colon cancer cells

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外科,湖北武汉430022 [2]华中科技大学同济医学院病原生物系微生物学教研室,湖北武汉430030

出　　处：《中国普通外科杂志》2009年第4期343-347,共5页China Journal of General Surgery

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：目的探讨survivin靶向SiRNA重组表达载体对人结直肠癌细胞生长的抑制作用和对5-氟尿嘧啶(5-FU)的化疗增敏作用。方法构建survivin靶向SiRNA重组表达载体并转染结肠癌细胞,通过RT-PCR和westernblot方法检测survivin的表达;采用MTT法和流式细胞仪检测SiRNA对不同浓度5-FU的化疗增敏作用。结果经酶切鉴定和测序结果证实survivin靶向SiRNA重组表达载体构建成功;其对结直肠癌细胞survivinmRNA和蛋白的表达抑制率分别为36.33%和44.65%,可以协同增强5-FU对结直肠癌细胞生长的抑制作用。结论survivin靶向SiRNA重组表达载体能有效抑制survivin基因表达,并能协同增强5-FU对结直肠癌细胞的杀伤作用和诱导凋亡作用。Objective To construct SiRNA recombinant expression vector targeting survivin gene and study its effect on inhibition of proliferation of human colon cancer cells and enhancement of chemotherapy sensitivity. Methods SiRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cells. The effect of SiRNA recombinant expression vector was detected by RT-PCR and western blot. Enhancement of chemotherapy sensitivity was observed by MTT reduction assay and flow cytometry. Results SiRNA recombinant expression vector targeting survivin gene was constructed successfully confirmed by restriction endonuclease and sequence analysis, and its inhibition ratio on survivin mRNA and protein levels was 36.33% and 44.65% , respectively. It could enhance the inhibitory effect of 5-Fu on growth of colon cancer cells. Conclusions The SiRNA recombinant expression vector targeting survivin gene can effectively inhibit the expression of survivin gene. It also can synergetically enhance the chemotherapy sensitivity and apoptosis induction of 5- Fu on human colon cancer cells.

关 键 词：结直肠肿瘤 SURVIVIN RNA干扰 基因治疗 化疗 

分 类 号：R730.5[医药卫生—肿瘤]