四氢异喹啉类化合物HZ08通过降低白血病细胞的葡萄糖神经酰胺合成酶和MDR1表达水平逆转耐药作用

HZ08 reverses multidrug resistance in leukemia cells by reducing glucosylce-ramide synthase and MDR1

通过对比K562/A02细胞株和耐药白血病临床样本的实验结果,探讨四氢异喹啉类化合物HZ08对多药耐药的逆转作用和逆转机制。将阿霉素与10μmol/L HZ08合用,MTT法测得阿霉素对临床耐药的白血病细胞的IC50为0.60μmol/L,对K562/A02细胞的IC50为0.83μmol/L,逆转倍数分别为27.87和20.49。使用免疫印迹法检测细胞内葡萄糖神经酰胺合成酶(GCS)表达水平,荧光实时定量聚合酶链反应检测MDR1的mRNA表达水平。结果显示:各浓度的HZ08(15,20,25μmol/L)与阿霉素合用均能降低GCS的表达,与单用阿霉素组相比,阿霉素与HZ08合用可显著降低MDR1的mRNA表达(P<0.01)至空白组水平。结果表明,HZ08与阿霉素合用给药,对K562/A02细胞株和耐药的临床耐药白血病细胞均有较强的逆转作用,其机制可能与降低细胞内GCS蛋白的表达和MDR1 mRNA的表达有关。

In this paper the influence of HZ08 on multidrug resistance and possible mechanism were explored.It was found that the IC50 of adriamycin in the presence of 10 μmol/L HZ08 in clinical sample cells was 0.60 μmol/L,while the IC50 in K562/A02 cells was 0.83 μmol/L by MTT.The reversal folds of MDR were 27.87 and 20.49,respectively.For further mechanism study of HZ08,the expression level of glucosylceramide synthase（GCS） protein in K562/A02 and resistant cells from clinical samples were analyzed by Western blot,while the mRNA of MDR1 was detected by real-time fluorescent quantitative polymerase chain reaction（FQ-RT-PCR）.Combined with adriamycin,all concentrations（15,20,25 μmol/L） of HZ08 decreased the expression of GCS protein.In addition,compared with the group of treatment with adriamycin alone,combined treatment of HZ08 and adriamycin decreased the expression of MDR1 significantly（P 0.01）.Therefore,HZ08 possibly reverses multidrug resistance by its joint action with adriamycin on GCS and MDR1 in both cell line and clinical sample cells.