四株人白血病多药耐药细胞系的建立及其生物学特性的初步研究

我们对人红白血病细胞株K562和人早幼粒细胞白血病细胞株HL－60应用高三尖杉酯破（HHT）或长春新碱（VCR），采用反复短期暴露法，并逐渐增加HHT和VCR的浓度，成功地建立了K562／VCR、K562／HHT、HL－60／VCR和HL－60／HHT四株具有高度耐药性的多药耐药（MDR）细胞系。免疫组织化学染色显示，K562／VCR、K562／HHT及HL60／VCR三株MDR细胞系有卜糖蛋白（Pgp）的高度表达，而HL－60／HHT细胞系没有pgP的表达。四株MDR细胞系均未见有多药耐药相关蛋白（MRP）的表达。5μg／ml异搏定和50μg／ml红霉素都能部分逆转K562／VCR、K562／HHT和HL－60／VCR三株MDR细胞系的耐药性，但其对HL－60／HHT细胞系的耐药性几乎无逆转作用。四株MDR细胞系与其相应敏感细胞系相比，前者对DNR的摄取均减少，外排均增加。2．5～10μg／ml异搏定能增加K562／VCR、K562／HHT和HL－60／VCR三株MDR细胞系对DNR的摄取量和滞留量，且其作用随异搏定浓度的增加而增强，但50～200μgn／ml红霉素不能增加它们对DNR的摄取量和滞留量。上述浓度异搏定或红霉素均不能增加HL－60／HHT细胞系对DNR的摄取量和滞留量。

A PRELIMINARY STUDY ON ESTABLISHMENT AND CHARACTERIZATION OF FOUR HUMAN MULTIDRUGRESISTANT LEUKEMIC CELL LINES

We established four human highly multidrug- resistant (MDR ) leukemic cell lines (K562/VCR, K562/HHT, HK-60/VCR and HL-60/HHT) from human erythroleukemiacell line K562 and human promyelocytic leukemia cell line HL-60 by short and exposures to HHT or VCR with gradually increasing concentration of HHT or VDR. Immunohistochemical staining demonstrated K562/VCR, K562/HHT and HL-60/VCR MDR cell lines overexpressed p-glycoprotein (Pgp), but HL-60/HHT cell line didn' t. No expression of MDR-related protein (MRP) was found in all four MDR cell lines. The resistance of K562/VCR or K562/HHT or HL-60/VCR MDR cell line could be partly reversed, hut that of HL-60/HHT MDR cell line could be hardly done by 5μg/ml verapamil or 50μg/ml erythromycin.Both DNR accumulation reduced and DNR efflux increased in four MDR cell lines as compared with their parental cell lines. 2. 5~10μg/ml verapamil was able to increase DNR accumulation and retention in K562/VCR, K562/HHT and HL-60/VCR MDR cell lines, and the more verapamil, the more DNR accumulation and retention, but not in HL-60/HHT MDR cell line. 50-200μg/ml erythromycin was unable to increase NDR accumulation and retention on four MDR cell lines.