抑制BCRP表达逆转肝细胞癌MDR的研究
Reverse multidrug resistance of hepatocellular carcinoma by suppressing breast cancer resistance protein gene
李涛 1曾贵利 2陈龙 2吴进峰 2陈勇 1余少红 3王晓波 3龚建平1
作者信息
- 1. 重庆医科大学附属第二医院 肝胆外科,重庆,400010
- 2. 重庆医科大学附属第二医院 肝病研究所,重庆,400010
- 3. 重庆市涪陵中心医院肝胆外科,重庆,400800
- 折叠
摘要
目的:研究小片段RNA干扰对肝癌耐药细胞系HEPG2/ADM中BCRP基因及其蛋白产物BCRP表达的抑制作用和逆转其耐药性的效果.方法:构建针对BCRP基因pSUPER-RNAi质粒,转染肝癌耐药细胞HEPG2/ADM.通过RT-PCR和Western Blot在mRNA和蛋白水平评价RNAi对BCRP表达的影响.MTT法检测RNAi逆转HEPG2/ADM细胞耐药性的效果,按实验组和对照组细胞的半数抑制剂量(IC50)计算RNAi对各种细胞耐药倍数的改变.结果:在耐药肝癌细胞HEPG2/ADM中,RNAi明显抑制了BCRP mRNA和蛋白产物BCRP的表达水平,其表达仅为对照组细胞的22.55%和37.49%(P<0.01).在相同浓度化疗药物的作用下,RNAi组HEPG2/ADM细胞耐药性显著下降,对ADM的耐药逆转倍数为3.55倍.结论:针对多药耐药基因BCRP,RNAi具有强大的逆转肝癌耐药细胞HepG2多药耐药的作用.为从基因水平逆转MDR展示了良好的前景.
Abstract
AIM: To investigate the suppression of breast cancer resistance protein ( BCRP) gene and protein by small interfering RNA ( siRNA) and the restoration of its sensitivity to chemothera-peutic drugs in multidrug-resistant hepatocellular carcinoma cell line HEPG2/ADM. METHODS: BCRP targeted siRNA duplexes were transfected into HEPG2/ADM. The suppression of BCRP gene and protein product was analyzed by RT-PCR and Western blot, respectively. MTT assay was performed to measure the reverse effect of BCRP-siRNA based on the results of IC50. RESULTS: The over-expression of BCRP gene was suppressed efficiently by siRNA. The levels of BCRP mRNA and protein expression in the transfected HEPG2/ADM cells reduced to 22.55% and 37.49% in comparison with those in the controls, respectively. Their resistance to ADM was reversed by 3.55 folds, which indicated the restoration of its sensitivity to drugs. CONCLUSION: The sequence specific RNAi inhibits BCRP mRNA and protein expression in the HepG2/ADM cells. It may reverse multidrug resistance phenotype and thus provide promising therapeutic modalities in the treatment of hepatocellular carcinoma (HCC).
关键词
抗药性,多药/乳腺癌耐药蛋白/RNA干扰/癌,肝细胞Key words
drug resistance/multiple/breast cancer resistance protein/RNA interference/carcinoma/hepatocellular引用本文复制引用
基金项目
国家自然科学基金(30801126)
重庆市卫生局资助项目(05-2-224)
出版年
2009
NETL
NSTL
维普
万方数据