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| RNAi抑制肾癌细胞MDR1基因表达及对顺铂化疗耐药的逆转 | |
| 引用本文： | 郝怡鑫,贺铮雯,杜楠,鲁琼,肖文华,沈茜.RNAi抑制肾癌细胞MDR1基因表达及对顺铂化疗耐药的逆转[J].肿瘤,2010,30(2). |
| 作者姓名： | 郝怡鑫 贺铮雯 杜楠 鲁琼 肖文华 沈茜 |
| 作者单位： | 1. 第二军医大学附属长海医院实验诊断科,上海,200433;解放军总医院第一附属医院肿瘤科,北京,100037 2. 第二军医大学附属长海医院实验诊断科,上海,200433 3. 解放军总医院第一附属医院肿瘤科,北京,100037 |
| 摘 要： | 目的:研究RNA干扰(RNA interference,RNAi)对肾癌细胞多药耐药基因1(multidrug resistance,MDR1)表达的抑制作用,并分析干扰前后肾癌细胞对顺铂(cisplatin)敏感度的变化.方法:根据MDR1基因序列设计3条小干扰RNA(small interfering RNA,siRNA),转染肾癌A498细胞;RT-PCR法检测转染前后MDR1 mRNA表达水平的变化,筛选出干扰效率最高的siRNA序列;进而利用慢病毒包装siRNA重组质粒,感染A498细胞,RT-PCR法筛选沉默效果最好的细胞株进行克隆;Western印迹法检测MDR1蛋白表达水平;MTT法检测干扰前后顺铂对半数细胞抑制浓度(half inhibitory concentration,IC_(50))的变化.结果:3条siRNA序列均能不同程度地抑制细胞MDR1基因的表达,其中siRNA-1序列能更有效地封闭MDR1基因,使MDR1 mRNA表达水平下降67%;筛选得到稳定转染的细胞株与未干扰细胞株相比,MDR1蛋白表达量明显下降(P<0.01),并使顺铂对细胞的IC_(50)值降低约83.37%(P<0.01).结论:RNAi能有效抑制肾癌A498细胞MDR1基因的表达,并可显著增加其对顺铂的敏感度,从而使肾癌细胞化疗耐药逆转成为可能. |
| 关 键 词： | 肾肿瘤 RNA 小分子干扰 基因 MDR 顺铂 A498细胞 |
Suppression of MDR1 gene expression and reversal of cisplatin resistance in renal carcinoma cells by RNA interference |
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| HAO Yi-xin,HE Zheng-wen,DU Nan,LU Qiong,XIAO Wen-hua,SHEN Qian.Suppression of MDR1 gene expression and reversal of cisplatin resistance in renal carcinoma cells by RNA interference[J].Tumor,2010,30(2). | |
| Authors: | HAO Yi-xin HE Zheng-wen DU Nan LU Qiong XIAO Wen-hua SHEN Qian |
| Institution: | HAO Yi-xin1,2,HE Zheng-wen1,DU Nan2,LU Qiong1,XIAO Wen-hua2,SHEN Qian1 (1.Department of Laboratory Diagnosis,Changhai Hospital,Second Military Medical University,Shanghai 200433,China,2.Department of Oncology,First Affiliated Hospital of People\'s Liberation Army General Hospital,Beijing 100037,China) |
| Abstract: | Objective:To investigate the inhibitory effect of RNA interference (RNAi) on the expression of multidrug resistance (MDR1) gene and analyze the altered sensitivities of human renal carcinoma cell line to cisplatin.Methods:Three small interfering RNA (siRNA) sequences targeted MDR1 gene were synthesized and transfected into renal carcinoma A498 cells. The expression level of MDRl mRNA was measured by RT-PCR to identify the most effective siRNA sequence. The recombinant plasmid was packed by lentivirus and tr... |
| Keywords: | Kidney neoplasms RNA small interfering Genes MDR Cisplatin A498 cells |
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