环孢菌素A联合汉防己甲素逆转人白血病耐药细胞株K562/A02细胞多药耐药的分子生物学机制  

Biomolecular Mechanisms of Cyclosporine A,Tetrandrine and Their Combination on the Reversion of Multidrug Resistance in Human Leukemia Cell Line

作　　者：陈宝安[1] 郭晶晶[1] 程坚[1] 高峰[1] 丁家华[1] 许文林[2] 沈惠玲[2] 孙新臣[3] 成红艳[3] 高冲[1] 孙耘玉[1] 王骏[1] 赵刚[1] 宋慧慧[1] 鲍文[1] 李国宏[1] 刘莉洁[1] 陈文姬[1] 王雪梅[4] 

机构地区：[1]东南大学附属中大医院血液科,南京210009 [2]江苏大学附属人民医院血液科 [3]东南大学附属中大医院肿瘤科 [4]南京东南大学生物电子学国家重点实验室

出　　处：《中国中西医结合杂志》2008年第11期1010-1013,共4页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.39970832);国家自然科学基金资助项目(No.30740062);江苏省科技计划资助项目(No.BS2004024);高等学校博士学科点专项科研基金(No.20070286042)

摘　　要：目的多药耐药(multidrug resistance,MDR)是导致肿瘤化疗失败的主要原因,本研究探讨环孢菌素A(cyclosporine A,CsA)及汉防己甲素(tetrandrine,Tet)联合应用对人白血病耐药细胞株K562/A02细胞MDR的逆转作用及其分子生物学机制。方法K562/A02细胞经CsA和Tet作用后,以流式细胞仪(FCM)检测细胞内柔红霉素(daunorubicin,DNR)浓度和P-糖蛋白(P-glycoprotein,P-gp)的表达,逆转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)检测多药耐药基因(multidrug resistancegene,mdr1)mRNA表达水平。结果K562/A02细胞内DNR浓度为K562的19%,CsA(1mg/L)及Tet(0.1mg/L)单独及联合作用K562/A02细胞48h后细胞内DNR的浓度分别为K562的60%、65%、98%。K562和K562/A02细胞P-gp荧光强度分别为0.18%和96.51%,K562/A02细胞经CsA(1mg/L)和Tet(0.1mg/L)单独及联合处理后细胞P-gp荧光强度分别为75.32%,76.86%和48.61%。K562/A02细胞经过两药单独作用后耐药株mdr1-mRNA下调微弱,联合用药下调效果明显。结论环孢菌素A和汉防己甲素均可逆转白血病细胞的耐药,且两药联合作用逆转效果增强,具有协同作用。Objective Multidrug resistance （MDR） plays an important role in the failure of chemotherapy on malignant tumors. This study was to investigate the biomolecular mechanisms of cyelosporine A （ CsA）, tetrandrine （Tet） and their combination on multidrug resistance in cell line K562/A02. Methods K562/A02 cells were treated with CsA and （or） Tet. The intracellular daunorubicin （DNR） concentration and the expression of P-glycoprotein （P-gp） were observed by flow eytometry assay. The mRNA expression of mudtiding resistance gene （ mdrl ） was measured by fluorescent semi-quantitative reverse transeriptase polymerase chain reaction （RT-PCR）. Results CsA and Tet （alone or combination） elevated the intracellular DNR concentration in K562/A02 cells （the fluorescence intensity of intracellular DNR in K562/A02 cells was 60% , 65% and 98% respectively of that in K562 cells）. The fluorescence intensity of P-gp in K562 and K562/A02 cells was 0.18% and 96.51%. The P-gp expression was down-regulated after treated with CsA, Tet and both （75.32% , 76.86% and 48.61% ） ; mdrl mRNA was also down-regulated, and the effect of their combination was more obvious. Conclusion MDR can be partially reversed by CsA or Tet, the combination of both drugs shows a great synergistic reversal effect.

关 键 词：环孢菌素A 汉防己甲素 K562/A02细胞 多药耐药 P-糖蛋白 

分 类 号：R733.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]