肝癌多药耐药细胞株的建立及其多药耐药机理的研究

王宝成; 郭军; 狄剑时; 师秋丽; 姚长樱

肝癌多药耐药细胞株的建立及其多药耐药机理的研究

Establishment of multidrug resistance cell strain of liver cancer and reserch of its multidrug resistance mechanism

摘要

摘要: 为建立人肝癌多药耐药细胞株并研究其多药耐药的机理，本文应用BEL－7402细胞株，通过不断提高培养液中阿霉素（Doxorubicin）的浓度，长期筛选培养，得到肝癌多药耐药株BEL7402/DoX。经MTT法检测BEL－7402对长春新碱（VCR）等8种抗癌药的抗性提高了27－1100倍不等。以流式细胞技术检测了此细胞株表面MDR1蛋白P－gp、多药耐药相关蛋白MRP及谷胱甘肽硫转移系统（GSH/GST）的表达；用RT－PCR方法检测了MDR及MRP基因表达水平。发现BEL7402/Dox细胞表面P－gp表达为93.5～97.4％；MRP的表达为84.7～90.2％；RT－PCR证实此细胞株中有MDR及MRPmRNA的高表达；未发现GSH/GST的表达升高。

Abstract: For serching establishing human liver cancer multidrug resistance cell strain and reserch its drug resistance mechanism,the BEL-7402 cells were cultured orubicin by progressively cell strain BEL-7402/Dox increase concentration method,and the drug with Dox-resistance were selected for a resistance increased of BEL-7402 cells to vincristinelong period screening cultural.The (VCR) and other 8 anticancer drugs 27-1100 times respectivly by MTT assay.The MDR1 expression product membrane efflux p-glycoprotein and multidrug resistanceassociated(MRP)as well as GSH/GST were examined,the MDR and MRP gene expression were also detected using RT-PCR.The results showed that p-gp level of BEL-7402/Dox were 93.5 to 97.4%MRP were 84.7 to 90.2%,the level of MDR and MRP mRNA expression were also relatively high,no over experssion of GSH/GST was found.

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