Reversal of multidrug resistance of hepatocellular carcinoma by siRNA/mdr1

潘光栋; 严律南; 王新平; 杨家印; 夏庆杰; 闫乃红; 陈清英; 张发强

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Reversal of multidrug resistance of hepatocellular carcinoma by siRNA/mdr1

更新时间：2026-01-09

- Reversal of multidrug resistance of hepatocellular carcinoma by siRNA/mdr1
- Journal of Army Medical University Vol. 30, Issue 1, Pages: 35-38(2008)
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  CLC： R394-33;R73-36;R735.7
- Published：2008
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潘光栋,严律南,王新平,杨家印,夏庆杰,闫乃红,陈清英,张发强.RNA干扰逆转肝细胞癌多药耐药[J].陆军军医大学学报(原第三军医大学学报),2008,30(01):35-38.

　PAN Guang-dong,YAN Lu-nan,WANG Xin-ping,YANG Jia-yin,XIA Qing-jie,YAN Nai-hong,CHEN Qing-ying,ZHANG Fa-qiang.Reversal of multidrug resistance of hepatocellular carcinoma by siRNA/mdr1[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(01):35-38.
潘光栋,严律南,王新平,杨家印,夏庆杰,闫乃红,陈清英,张发强.RNA干扰逆转肝细胞癌多药耐药[J].陆军军医大学学报(原第三军医大学学报),2008,30(01):35-38. DOI：

　PAN Guang-dong,YAN Lu-nan,WANG Xin-ping,YANG Jia-yin,XIA Qing-jie,YAN Nai-hong,CHEN Qing-ying,ZHANG Fa-qiang.Reversal of multidrug resistance of hepatocellular carcinoma by siRNA/mdr1[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(01):35-38. DOI：

摘要

目的筛选高效dsRNA/mdr1，以备研究RNA干扰逆转肝细胞癌多药耐药之用。方法首先根据siRNA设计原则，以多药耐药基因（mdr1）为靶基因，设计并选择4～5条siRNA/mdr1，经BLAST后体外转录法合成dsRNA/mdr1，用Oligofectamine试剂分别转染HepG2/mdr1，然后从mRNA、蛋白（P-gp）表达水平和细胞功能变化评价HepG2/mdr1耐药性被逆转的程度，比较各个dsRNA/mdr1的逆转效率，筛选出有效的siRNA/mdr1。结果成功合成5条dsRNA/mdr1(其中1条为阴性对照)，dsRNA/mdr1-4 mRNA表达（18.73±1.33）％、蛋白表达变化(79.1±1.6)%~（16.8±0.4)%与其他各组细胞比较，有显著性差异；细胞内柔红霉素(DNR)累积量也较其他组明显增加（平均荧光强度79.58，阳性率84.25%，P0.05）。结论体外转录法结合脂质体转染适用于筛选高效siRNA，肯定了siRNA干扰序列能够有效阻抑mdr1基因编码蛋白p170的功能。

Abstract

Objective To screen effective dsRNA/mdr1 for studying the reversal of multidrug resistance of hepatocellular carcinoma (HCC) by RNA interference (RNAi). Methods dsRNA/mdr1 targeting multidrug resistance gene (mdr1) was designed and synthesized by in vitro transcription. HepG2/mdr1 in 6 groups was transfected with the complex of different dsRNA/mdr1 (dsRNA/mdr1-1

dsRNA/mdr1-2

dsRNA/mdr1-3

dsRNA/mdr1-4

dsRNA/mdr1-5) using Oligofectamine. Then the cells were collected to measure the expression of mRNA/mdr1

P-glycoprotein and the accumulation of DNR. Results Five dsRNA/mdr1 were successfully synthesized including one negative. The expression of mRNA/mdr1 [(18.73±1.33)％] and P-glycoprotein [from (79.1±1.6)% before transfection down to (16.8±0.4)%] in dsRNA/mdr1-4 was significantly lower than that of other groups (P0.05). Accumulation of DNR in dsRNA/mdr1-4 was higher than that in other groups (P0.05). Conclusion In vitro transcription combined with liposome is fit to screen the effective dsRNA/mdr1. siRNA/mdr1 could suppress the expression of P-glycoprotein coded by mdr1 gene.

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