环孢菌素D衍生物PSC833逆转K562/A02细胞多药耐药的研究  

Reversal of multidrug-resistance in human leukemia cell line K562/A02 by a cyclosporin D analogue PSC 833

作　　者：戴辉[1] 罗绍凯[1] 尹爱华[2] 彭爱华[1] 

机构地区：[1]中山医科大学附属第一医院血液科 [2]中山医科大学达安基因诊断中心

出　　处：《中华血液学杂志》2002年第1期23-26,共4页Chinese Journal of Hematology

摘　　要：目的　证实PSC 833逆转剂的高效性 ,探讨PSC 833逆转肿瘤细胞多药耐药的机制。方法　以人红白血病细胞系K5 6 2及其耐药细胞系K5 6 2 /A0 2 (耐阿霉素 )为实验研究对象 ,采用MTT法检测细胞毒性 ;直接免疫荧光测定法检测P糖蛋白 (P gp)表达水平 ;RT PCR法检测mdr1mRNA水平 ;以流式细胞术测定两种细胞系内柔红霉素 (DNR)的潴留来反映P gp的外排功能。结果　与K5 6 2细胞系相比 ,K5 6 2 /A0 2耐药细胞系mdr1mRNA及P gp高表达 ,DNR潴留减少。 1μmol/L的PSC 833对K5 6 2 /A0 2细胞的mdr1mRNA及P gp表达水平无明显影响 (P >0 .0 5 ) ,PSC 833对K5 6 2 /A0 2细胞的DNR细胞毒性有剂量依赖性增敏作用 ,其增敏作用至少是环孢菌素A(CsA)、维拉帕米 (Ver)的 3倍。PSC 833能增加K5 6 2 /A0 2耐药细胞系的DNR潴留。 1μmol/L的PSC 833能使K5 6 2 /A0 2细胞内DNR潴留量恢复至K5 6 2细胞的 10 0 .9% ,而 10 μmol/LCsA只恢复至K5 6 2细胞的 86 .9% ,PSC 833对K5 6 2细胞系的DNR细胞毒性及DNR潴留均无明显影响 (P >0 .0 5 )。结论　PSC 833较CsA、Ver逆转活性至少高 3～10倍 ,其逆转K5 6 2 /A0 2多药耐药的机制可能是通过抑制P gp功能 ,而非直接下调mdr1mRNA及PObjective To explore the efficacy of PSC 833 on multidrug resistance (MDR) reversal and its mechanism. Methods Human erythroleukemic cell line K562 and its doxorubicin resistant counterpart K562/A02 were used in the study. Cytotoxicity was assessed by MTT assay, P gp expression by direct immunofluorescence and mdr1 mRNA expression by reverse transcriptase polymerase chain reaction (RT PCR) with β actin as internal control. Intracellular DNR retention was measured with flow cytometry. Results K562/A02 cells displayed high levels of mdr1 mRNA and P glycoprotein and reduced DNR retention compared to their parental K562 cells. 1 μmol/L of PSC 833 had no effect on the levels of mdr1 mRNA and P gp expression in K562/A02 cells (P>0.05). PSC 833 conferred a dose dependent increase on chemosensitivity of K562/A02 to DNR, and its effect was at least 3 fold more potent than that of CsA or Ver. PSC 833 could increase DNR retention in K562/A02 cells. A 100.9% restoration of intracellular DNR retention of the level of K562 cells was gained by PSC 833 at 1.0 μmol/L in K562/A02 cells, whereas only a 86.9% restoration of DNR retention was obtained by CsA at 10 μmol/L in the K562/A02 cells. No effect on DNR sensitivity and retention was found in K562 cells (P>0.05). Conclusion PSC 833 is at least 3～10 fold more potent than CsA or Ver with respect to MDR reversing activity, and it may function by inhibiting the function of P gp and not reducing the levels of mdr1 mRNA and P gp directly.

关 键 词：环孢菌素类 多药抗药性 P糖蛋白 白血病 K562细胞系 

分 类 号：R733.7[医药卫生—肿瘤]