多药耐药相关蛋白在肝癌多药耐药细胞系HePG2／ADM中的作用  

The role of multi-drug resistance related proteins in hepatocellular carcinoma multidrug resistance cell line HePG2/ADM

作　　者：李高鹏[1] 陈孝平[1] 王其[1] 徐宗全[1] 张万广[1] 叶露[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科中心,武汉430030

出　　处：《中华普通外科杂志》2006年第11期766-768,共3页Chinese Journal of General Surgery

基　　金：卫生部临床学科重点项目(卫规财发[2001]321号)

摘　　要：目的研究4种耐药蛋白:多药耐药蛋白(multi-drug resistance protein 1,MDR1),多药耐药相关蛋白1(multi-drug resistance related protein 1,MRP1),肺耐药蛋白(lung resistance protein, LRP),乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)在肝癌多药耐药中的作用。方法通过培养液中阿霉素(adriamycin,ADM)浓度递增诱导法筛选培养,建立HePG2／ADM肝癌多药耐药细胞株;采用real time荧光定量PCR检测四种耐药蛋白mRNA在正常肝细胞系L02、肝癌细胞系HePG2及HePG2／ADM中的表达差异;Western blot检测3种细胞系中4种耐药蛋白的表达。结果(1)建立HePG2／ADM细胞系。MTT法检测阿霉素对HePG2／ADM细胞IC50为亲本细胞的282倍(P<0．05)。(2)HePG2／ADM中MDR1和BCRP mRNA表达分别是HePG2的400倍(F=87．49, P<0．05)和9倍(F=1006,P<0．05)。MRP1和LRP mRNA表达差异无统计学意义(FMRP1=3．43, FLRP=2．44,Pall>0．05)。(3)L02和HePG2中4种耐药蛋白mRNA的表达均无差异(FMDR1=1006, FBCRP=87．49,FMRP1=3．43,FLRP=2．44,Pall>0．05)。(4)Western blot检测发现HePG2／ADM细胞中MDR1,BCRP,LRP蛋白表达显著高于HePG2和L02细胞(FMDR1=28．68,FBCRP=18．60,FLRP= 6．28,Pall<0．05),MRPI蛋白表达不增高(FMRP1=0．70,P>0．05)。(5)4种耐药蛋白表达在HePG2和L02细胞差异均无统计学意义(FMDR1=28．68,FBCRP=18．60,FMRP1=0．70,FLRP=6．28, Pall>0．05)。结论MDR1和BCRP在HePG2／ADM多药耐药中起重要作用,HePG2／ADM多药耐药与MRP1和LRP可能无关。Objective To explore the roles of MDR1 （ multi-drug resistance protein 1 ）, MRP1 （ multi-drug resistance related protein 1 ）, LRP（ lung resistance protein） and BCRP（ breast cancer resistance protein） in the phenomenon of multi-drug resistance （MDR） in hepatoma. Methods Through exposing HePG2 cell line to gradually increased concentration of adriamycin （ ADM ） , HePG2 muhidrug resistant subcell line （HePG2/ADM） was induced. The MDR index of HePG2/ADM was investigated by the method of MTT. The expression of the four MDR proteins in the three cell lines （ L02, HePG2, HePG2/ADM） were investigated at mRNA and protein levels by real-time RT-PCR and Western blot respectively. Result The IC50 of the HePG2/ADM to ADM was 282 times as higher than HePG2 （ P 〈 0. 05 ）. The expression of MDR1 and BCRP mRNA in HePG2/ADM cells were respectively 400 （ F = 87.49, P 〈 0. 05 ） and 9 times higher than HePG2 cells （ F = 1006, P 〈 0. 05 ）, while the mRNA expressions of MRP! and LRP did not change significantly （ FMRPI= 3. 43, FLRP = 2. 44, Pall 〉 0. 05 ）. There was no difference between HePG2 and L02 cells for mRNA expressions of the four genes （ FMDRI = 1006, FBCRP = 87.49, FMRPI = 3. 43, FLRP = 2.44, Pall 〉0. 05）. At the protein level, the expressions of MDR1, BCRP and LRP in HePG2/ADM were significantly higher than HePG2 and L02 （ FMDRI ： 28. 68, FBCRP = 18. 60, FLRP = 6. 28, P 〈 0. 05 ）. In HePG2 and L02, there was no difference for the expressions of the four genes at the protein level （ FMDRI = 28.68, FBCRP = 18.60, FMRPI = 0. 70, FLRP = 6.28, Pall 〉 0. 05 ）. Conclusion The four genes are probably the normally expressed genes in the liver,The expression of MDR1 and BCRP could be upregulated by anticancer agents in vitro ,MDR1 and BCRP may play very important roles in the MDR of HePG2/ADM .However,MRPI and LRP may have no relations with the multi-drog resistaniec of HePG2/ADM cell lines.

关 键 词：癌 肝细胞 多药耐药相关蛋白类 抗肿瘤联合化疗方案 

分 类 号：R735.7[医药卫生—肿瘤]