| 逆转录病毒载体介导的RNAi抑制Bmi-1基因表达对MCF-7细胞的影响 |
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| 引用本文： | 武向梅,卜友泉,宋方洲,刘涛,张琴,袁成福,袁飞,秦琴.逆转录病毒载体介导的RNAi抑制Bmi-1基因表达对MCF-7细胞的影响[J].第四军医大学学报,2009(13). |
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| 作者姓名： | 武向梅 卜友泉 宋方洲 刘涛 张琴 袁成福 袁飞 秦琴 |
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| 作者单位： | 重庆医科大学重庆市分子医学与肿瘤研究中心;重庆医科大学生物化学与分子生物学教研室; |
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| 摘 要： | 目的:研究RNA干扰(RNAi)抑制Bmi-1基因表达后对乳腺癌细胞株MCF-7凋亡和增殖的影响.方法:将表达Bmi-1短发夹RNA(shRNA)的重组逆转录病毒载体pSuper-retro/Bmi-1si瞬时转染MCF-7乳腺癌细胞株,携带针对绿色荧光蛋白干扰序列的载体pSuper-retro/GFPsi作为阴性对照.用RT-PCR和Western Blot分别从mRNA,蛋白水平检测Bmi-1的表达;MTT法检测细胞增殖;流式细胞术检测MCF-7细胞周期和凋亡的变化.结果:pSuper-retro/Bmi-1si明显抑制MCF-7细胞Bmi-1 mRNA及蛋白表达.与空白对照组及阴性对照组相比较凋亡细胞数无明显增加;抑制Bmi-1基因表达使MCF-7细胞G1/S周期转换及细胞增殖速率受抑制.结论:表达shRNA的重组逆转录病毒载体pSuper-retro/Bmi-1si能显著抑制乳腺癌细胞株MCF-7中Bmi-1基因表达,从而有效抑制MCF-7细胞增殖,为乳腺癌的基因治疗提供了实验依据.
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| 关 键 词： | Bmi-1基因 重组逆转录病毒 RNA干扰 |
Effect of retrovirally mediated RNA interference targeting human Bmi-1 gene on MCF-7 cell line |
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| WU Xiang-Mei,BU You-Quan,SONG Fang-Zhou,LIU Tao,ZHANG Qin,YUAN Chen-Fu,YUAN Fei,QIN Qin,Chongqing Research Center of Molecular Medicine , Tumor.Effect of retrovirally mediated RNA interference targeting human Bmi-1 gene on MCF-7 cell line[J].Journal of the Fourth Military Medical University,2009(13). |
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| Authors: | WU Xiang-Mei BU You-Quan SONG Fang-Zhou LIU Tao ZHANG Qin YUAN Chen-Fu YUAN Fei QIN Qin Chongqing Research Center of Molecular Medicine Tumor |
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| Institution: | WU Xiang-Mei1,2,BU You-Quan1,SONG Fang-Zhou1,LIU Tao1,ZHANG Qin1,YUAN Chen-Fu1,YUAN Fei1,QIN Qin1,21Chongqing Research Center of Molecular Medicine and Tumor,2Department of Biochemistry and Molecular Biology,Chongqing Medical University,Chongqing 400016,China |
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| Abstract: | AIM:To investigate the inhibition effects of Bmi-1(B-cell-specific moloney murine leukemia virus insertion site 1) with RNA interference(RNAi) on the apoptosis and proliferation of breast carcinoma MCF-7 cells. METHODS: Recombinant retroviral Bmi-1 gene short hairpin RNA(shRNA) expression vector pSuper-retro/Bmi-1 si was transiently transfected into breast carcinoma MCF-7 cells and pSuper-retro/GFP si vector was used as negative control. Bmi-1 mRNA and protein of the transfected cells were examined with rev... |
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| Keywords: | Bmi-1 gene recombinant retrovirus RNA interference |
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