Subcellular daunorubicin distribution and its relation to multidrug resistance phenotype in drug-resistant cell line SMMC-7721/R

作     者：Jia-Yin Yang Hua-You Luo Qi-Yuan Lin Zi-Ming Liu Lu-Nan Yan Department of General Surgery,West China Hospital,Sichuan University,Chengdu 610044,Sichuan Province,China Ping Lin Cancer Research Institution,West China Hospital,Sichuan University,610044,Sichuan Province,China Jie Zhang Department of Confocal Laser Scanning Microscopy,West China Hospital,Sichuan University,610044,Sichuan Province,China Shong Lei Cancer Biotechnological Treatment Center,West China Hospital,Sichuan University,610044,Sichuan Province,China 

作者机构：Department of General surgery First Affiliated Hospital Zhejiang University College of Medicine Hangzhou 310003 China. yjy7429@*** 

出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)

年 卷 期：2002年第8卷第4期

页      面：644-649页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：the grant from National Nature Science Foundation of China No.39770723 

主　　题：柔红素 SMMC-7721/R细胞 多药耐药 发生机制 RT-PCR 

摘      要：AIM: To investigate the correlation between subcellulardaunorubicin distribution and the multidrug resistancephenotype in drug-resistant cell line SMMC-7721/***: The multidrug resistant cell line SMMC-7721/R, a human hepatocellular carcinoma cell line,was established. Antisense oligonudeotides (AS-ODN)were used to obtain different multidrug resistancephenotypes by inhibiting the expression of mdr1 geneand/or multidrug resistance-related protein gene(mrp)using Lipofectamine as delivery agent. Expression ofmdr1 and mrp genes was evaluated by RT-PCR andWestern blotting. Intracellular daunorubicin (DNR)concentration was measured by flow *** DNR distribution was analyzed by confocallaser scanning microscopy. Adriamycin (ADM) and DMRsensitivity was examined by MTT ***: Low level expression of mdr1 and mrp mRNAsand no expression of P-Glycoprotein(P-gp) andmultidrug resistance-related protein (P190) weredetected in parental sensitive cells SMMC-7721/S, butover-expression of these two genes was observed indrug-resistant cell SMMC-7721/R. The expression ofmdr1 and mrp genes in SMMC-7721/R cells was downregulated to the level in the SMMC-7721/S cells by AS-ODN. Intracellular DNR concentration in SMMC-7721/S cells was 10 times higher than that in SMMC-7721/Rcells. In SMMC7721/S cells intracellular DNRdistributed evenly in the nucleus and cytoplasm, whilein SMMC-7721/R cells DNR distributed in a punctatepattern in the cytoplasm and was reduced in thenucleus. DNR concentration in SMMC-7721/R cells co-transfected with AS-ODNs targeting to mdr1 and mrpmRNAs recovered to 25 percent of that in SMMC7721/Scells. Intracellular DNR distribution pattern in drug-resistant cells treated by AS-ODN was similar to drug-sensitive cell, and the cells resistance index (RI) to DNRand ADM decreased at most from 88.0 and 116.0 to 4.0and 2.3, respectively. Co-Transfection of two AS-ODNsshowed a stronger synergistic effect than ***: P-gp