[1]潘光栋,严律南,王新平,等.RNA干扰逆转肝细胞癌多药耐药[J].第三军医大学学报,2008,30(01):35-38.
 PAN Guang-dong,YAN Lu-nan,WANG Xin-ping,et al.Reversal of multidrug resistance of hepatocellular carcinoma by siRNA/mdr1[J].J Third Mil Med Univ,2008,30(01):35-38.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第01期
页码:
35-38
栏目:
论著
出版日期:
2008-01-15

文章信息/Info

Title:
Reversal of multidrug resistance of hepatocellular carcinoma by siRNA/mdr1
作者:
潘光栋严律南王新平杨家印夏庆杰闫乃红陈清英张发强
广西医科大学第五附属医院肝胆外科;四川大学华西医院普通外科
Author(s):
PAN Guang-dong YAN Lu-nan WANG Xin-ping YANG Jia-yin XIA Qing-jie YAN Nai-hong CHEN Qing-ying ZHANG Fa-qiang
Department of Hepatobiliary Surgery, the Fifth Affiliated Hospital, Guangxi Medical University;Department of General Surgery, West China Hospital, Sichuan University
关键词:
肝细胞癌多药耐药RNA干扰
Keywords:
hepatocellular carcinoma multidrug resistance RNA interference
分类号:
R394-33; R73-36; R735.7
文献标志码:
A
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%,P<0.05)。  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 (P<0.05). Accumulation of DNR in dsRNA/mdr1-4 was higher than that in other groups (P<0.05).   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.

参考文献/References:

潘光栋, 严律南, 王新平, 等. RNA干扰逆转肝细胞癌多药耐药[J].第三军医大学学报,2008,30(1):35-38.

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更新日期/Last Update: 2008-05-05