三唑类抗真菌药联合多柔比星逆转白血病细胞株耐药性研究

目的探讨三唑类抗真菌药伊曲康唑、氟康唑联合多柔比星逆转白血病细胞株耐药的作用。方法伊曲康唑、氟康唑或PSC833（阳性对照）分别联合多柔比星与人类慢性粒细胞白血病红白血病耐多柔比星细胞株K562／ADR共同培养，采用CCK-8法检测K562／ADR细胞增殖，流式细胞术检测K562／ADR细胞内多柔比星平均荧光强度，蛋白印迹法检测K562／ADR细胞7H2AX的表达。结果1μg／ml伊曲康唑及0．5μg／mlPSC833可将K562／ADR对多柔比星的J岛从38．30μg／ml降至8．59μg／ml和24．64μg／ml，且呈剂量依赖性。1Ng／ml伊曲康唑或0．5μg／mlPSC833联合多柔比星作用K562／ADR细胞3h和6h后，细胞内多柔比星平均荧光强度相对单加多柔比星组增加了1．54倍（3h）、1．50倍（6h）或5．97倍（3h）、5．83倍（6h）。伊曲康唑或PSC833联合多柔比星可显著增加K562／ADR细胞7H2AX的表达。结论伊曲康唑通过提高细胞内多柔比星浓度及协同增强细胞DNA的损伤，恢复K562／ADR对多柔比星的敏感性，而氟康唑则无作用。

Reversal effect of triazole antifungal combined with adriamycin on multidrug resistance in leukemia cells

Objective To investigate the reversal effect of triazole antifungal itraconazole, fluconazole combined with adriamycin on multidrug resistance in leukemia cells. Methods Human chronic myelogenous leukemia adriamycin resistant cell lines K562/ADR cells were incubated with itraconazole, fluconazole, or PSC833 （positive control） combined respectively with adriamyein. CCK-8 assay was used to assess cell proliferation of K562/ADR. The mean fluorescence intensity of intracellular ADR was measured by flow cytometry. The marker of DNA damage ＂yH2AX was detected by Western blot. Results 1μg/ml itraconazole and 0.5 μg/ml PSC833 can decrease K562/ADR IC50 of adriamycin from 38.30 μg/ml to 8.59 μg/ml and 24.64μg/ml in a dose-dependent manner. K562/ADR cells were incubated with 1 μg/ml itraconazole or 0.5 μg/ml PSC833 combined respectively with adriamycin for 3 h and 6 h, the mean fluorescence intensity of intracellular ADR were increased 1.54-fold （3 h）, 1.50-fold （6 h） or 5.97-fold （3 h）, 5.83-fold （6 h）. Itraconazole or PSC833 combined with adriamycin significantly increase the expression of ~H2AX in K562/ ADR cells. Conclusion Itraconazole can recover adriamyein sensitivity of K562/ADR by increasing the concentration of intracelullar adriamycin and synergistically increasing DNA damage, but not for fluconazole.