【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。更多还原

【Abstract】 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 ( MOR) in hepatoma. Methods Through exposing HePG2 cell line to gradually increased concentration of adriamycin ( ADM) , HePG2 multidrug 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 MRP1 and LRP did not change significantly (FMRP1= 3.43, FLRP= 2. 44, Pall> 0. 05). There was no difference between HePG2 and L02 cells for mRNA expressions of the four genes (FMDR1 = 1006, FBCRP= 87. 49, FMRP1= 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 ( FMDR1= 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 (FMDR1= 28.68, FBCRP= 18.60, FMRP1=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 up-regulated by anticancer agents in vitro. MDR1 and BCRP may play very important roles in the MDR of HePG2/ADM. However, MRP1 and LRP may have no relations with the multi-drug resistance of HePG2/ ADM cell lines.更多还原