靶向抑制肝癌耐药细胞多药耐药基因表达的实验研究 |
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引用本文: | 胡礼仪,张有顺,周业庭,臧家兰,吴柏华.靶向抑制肝癌耐药细胞多药耐药基因表达的实验研究[J].牡丹江医学院学报,2010,31(5):31-34. |
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作者姓名: | 胡礼仪 张有顺 周业庭 臧家兰 吴柏华 |
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作者单位: | 1. 沭阳人民医院检验科,江苏,沭阳,223600 2. 湖北医药学院附属东风医院肝脏外科研究所,湖北,十堰,442008 |
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摘 要: | 目的:构建含多药耐药基因MDR1的短发夹状RNA(shorthairp in RNA,shRNA)表达质粒,并观察其对肝癌耐药细胞Bel-7402/R的MDR1 mRNA的抑制作用。方法:利用分子克隆技术,将含MDR1的双链DNA,与经双酶切后的载体PGE-1连接,构建pshRNA-MDR1重组质粒,在脂质体的介导下转染肝癌耐药细胞株BEL-7402/R,RT-PCR分析MDR1mRNA的表达,流式细胞仪检测细胞P-gp的表达。结果:PCR和DNA测序证实了表达质粒构建成功,并能明显地抑制MDR1mRNA的表达;P-gp的表达阳性率降低了57.3%,P〈0.05。结论:构建的pshRNA-MDR1表达质粒能靶向抵制肝癌耐药细胞MDR1mRNA和P-gp的表达。
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关 键 词: | 基因 多药耐药 RNA干涉 肝癌 |
INHIBITION OF MDR1 EXPRESSION IN HEPATOCELLULAR CARCINOMA CELL BEL-7402/R BY VECTOR-MEDIATED RNA INTERFERENCE |
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Institution: | Hu Liyi et al.(Department of Clinical Laboratories,the People′s Hospital of Shuyang,Shuyang 223600) |
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Abstract: | Objective:To construct a recombinant plasmids generating short hairpin RNA contain of multi-drug resistance gene MDR1 in mammalian cells in order to investigate suppression of MDR1 mRNA in hepatocellular carcinoma cells Bel-7402/R.Methods: Using molecular cloning technique,double-stranded DNA which contained MDR1 cloned into PGE-1 vector digesting by two restricted endoenzymes according to its special orientation.Human hepatocellular carcinoma cells Bel-7402/R was transfected with recombinant plasmid by LyoVecTM,MDR1 mRNA was assessed by RT-PCR,we determined the expression of P-gp by flow cytometry(FCM).Results: The size of the PCR product was 669bp.DNA sequencing showed the sequence of recombinant vector pshRNA-MDR1 was successfully constructed,and suppressed MDR1 mRNA expression;positive rate of P-gp receded from 77.7% to 20.4%,P0.05.Conclusion: The result showed that the recombinant plasmid constructed by the authors can suppress the expression of MDR1 mRNA and P-gp in hepatocellular carcinoma cells Bel-7402/R. |
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Keywords: | gene MDR RNA interference hepatocellular carcinoma |
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