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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) is the main cause of tumor chemotherapy failure. This study was to investigate the effect of combined use of cyclosporine A (CsA) and tetrandrine (Tet) on human leukemia Reversal effect of MDR on drug - resistant cell line K562 / A02 and its molecular biological mechanism. Methods After treated with CsA and Tet, the expression of daunorubicin (DNR) and P-glycoprotein (P-glycoprotein) in K562 / A02 cells were detected by flow cytometry (FCM) Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of multidrug resistance gene (mdr1). Results The DNR concentration in K562 / A02 cells was 19% of that in K562 cells. The intracellular DNR concentrations of K562 / A02 cells treated with CsA (1 mg / L) and Tet (0.1 mg / L) alone or in combination with K562 / A02 cells were 60% 65%, 98%. The P-gp fluorescence intensity of K562 and K562 / A02 cells were 0.18% and 96.51%, respectively. The fluorescence intensities of P-gp in K562 / A02 cells treated with CsA (1 mg / L) and Tet 75.32%, 76.86% and 48.61% respectively. After K562 / A02 cells were treated with two drugs alone, the mdr1-mRNA of drug-resistant strains had a weak down-regulation, and the combination therapy significantly reduced the effect. Conclusion Cyclosporine A and tetrandrine can both reverse the drug resistance of leukemia cells, and the synergistic effect of synergistic effect between the two drugs is enhanced.