靶向bmi-1真核表达载体的构建及对SW480细胞的增殖研究  

Construction of Eukaryotic Expression Vector Targeting Bmi-1 and its Influences on Proliferation of SW480 Cells

作　　者：汪云霞[1] 魏亚明[2] 王晓华[1] 朱晓莹[1] 

机构地区：[1]广州医学院生化教研室,广东广州510182 [2]广州市第一人民医院中心实验室,广东广州510182

出　　处：《现代生物医学进展》2009年第12期2270-2272,2312,共4页Progress in Modern Biomedicine

基　　金：广州市科技计划项目(2007J1-C0091)

摘　　要：目的:构建针对原癌基因bmi-1的shRNA逆转录病毒表达载体,检测它对大肠癌细胞株SW480增殖的影响。方法:利用载体介导的RNA干扰技术,设计、合成靶向bmi-1基因、编码短发夹RNA的寡核苷酸序列,并设计无义对照序列,构建重组载体pSIREN-bmi-1及pSIREN-bmi-1-c,载体经脂质体2000介导转染SW480细胞,经MTT检测,观察bmi-1表达受抑后对细胞增殖的影响。结果:成功构建了针对bmi-1基因的shRNA表达载体。结论:针对bmi-1基因的shRNA表达载体能够明显抑制结肠癌细胞SW480的增殖能力。Objective： To construct Retrovirus-mediated shRNA expression vector targeting bmi-1, which have been transfected SW480 cells so as to investigate the proliferation effect of bmi-1 shRNA on SW480 cells. Methods： Through the mediated vector, we de- signed and constructed the complementary oligonucleotides encoding hairpin RNA which specifically targets bmi-1 gene, and the non- sense oligonucleotides were designed as control. Then, we transfected them into colon cancer SW480 cells by lipofectamineTM 200.0. The proliferation of SW480 cells was observed after the inhibition of bmi-1 by MTT. Results： The vector expressing shRNA targeting bmi-1_was constructed successfully. Conclusion： The vector expressing shRNA targeting bmi-1 can inhibit cellular proliferation.

关 键 词：BMI-1基因 RNA干扰 结肠癌细胞 转染 短发夹RNA 

分 类 号：Q78[生物学—分子生物学]