[1]杨毅,姜习新,王继见,等.RNA干扰MRP2表达逆转结肠癌细胞对长春新碱的耐药性[J].陆军军医大学学报(原第三军医大学学报),2008,30(12):1179-1182.
 YANG Yi,JIANG Xi-xin,WANG Ji-jian,et al.RNAi silenced MRP2 reverses drug resistance of colorectal cancer cell line to vinblastine[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(12):1179-1182.
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RNA干扰MRP2表达逆转结肠癌细胞对长春新碱的耐药性(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第12期
页码:
1179-1182
栏目:
论著
出版日期:
2008-06-30

文章信息/Info

Title:
RNAi silenced MRP2 reverses drug resistance of colorectal cancer cell line to vinblastine
作者:
杨毅姜习新王继见周洪伟
重庆医科大学:附属第二医院普通外科,临床检验诊断学省部共建教育部重点实验室,重庆市重点实验室
Author(s):
YANG Yi JIANG Xi-xin WANG Ji-jian ZHOU Hong-wei
Department of General Surgery, Second Affiliated Hospital, Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400010, China
关键词:
多药耐药MRP2RNA干扰质粒
Keywords:
multidrug resistance MRP2 RNAi plasmid
分类号:
R73-361;R730.23;R735.35
文献标志码:
A
摘要:
目的  构建MRP2基因的真核表达载体pGenSil-1-MRP2-siRNA,并观察其转染人结肠癌耐长春新碱细胞株HCT-8/V前后MRP2基因和ABCC2蛋白表达变化以及对化疗药物敏感性变化。  方法  设计MRP2特异性siRNA的序列,克隆至pGenSil-1质粒中,测序鉴定后,用脂质体将重组质粒转染至HCT-8/V细胞中,用定量RT-PCR和Western blot检测MRP2的表达,MTT实验测定pGenSil-1-MRP2-siRNA和长春新碱对HCT-8/V细胞增殖抑制作用。   结果  在结肠癌耐药细胞HCT-8/V中,RNAi明显抑制了MRP2 mRNA及蛋白的表达,在相同浓度化疗药物的作用下,RNA干扰组细胞凋亡比例显著高于对照组(P<0.01),表明细胞耐药性显著下降,对药物敏感性显著增高。  结论  成功构建了MRP2的siRNA真核表达载体,并且对结肠癌耐药细胞MRP2的表达有明显抑制作用,取得良好的逆转耐药效果。
Abstract:
Objective    To construct the RNAi eukaryotic vector of inhibitory member of MRP family (MRP2) gene and after the vector transfection into HCT-8/V cell line to observe its interfering effect on MRP2 expression and the changed sensibility to vinblastine.     Methods    The specific siRNA sequence was designed according to the MRP2 sequence. The sequence was cloned into pGenSil-1 and then sequenced. The recombinant plasmid was transfected into HCT-8/V cells by liposome. Then MRP2 expression in the transfected cells was analyzed by FQ-PCR and Western blotting. MTT test was used to measure the inhibitory effects of pGenSil-1-MRP2-siRNA on the proliferation of HCT-8/V cells treated with vinblastine.      Results    The overexpression of MRP2 was suppressed efficiently by the introduction of small interfering RNA that caused sequence-specific gene silence. The apoptotic rate of HCT-8/V cells transfected with pGenSil-1-MRP2-siRNA increased significantly when the cells were exposed to vinblastine as compared with that of nontransfected cells (P<0.01), indicating the drug resistance of the transfected cells decreased.     Conclusion    The RNAi eukaryotic vector pGenSil-1-MRP2-siRNA is constructed successfully. The RNAi we constructed inhibits MRP2 expression effectively and reverses the drug resistance of HCT-8/V cells.

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更新日期/Last Update: 2008-06-16