长期传代后HL-60细胞内5’-核苷酸酶-n基因表达与阿糖胞苷耐药性的关系

Cytosolic 5'-nucleotidase-n gene expression and Ara-C drug resistance in long-term subcultured HL-60 cells

关键词:  阿糖胞苷  人早幼粒白血病细胞系  胞浆5’-核苷酸酶-Ⅱ  基因表达

Key words:  cytosine arabinoside  human leukemia HL-60 cells  cytosolic 5'-lucleotidase H  gene expression

摘要:

目的观察白血病细胞株(HL-60)及其耐药细胞株(HL-60/R)胞浆5’-核苷酸酶-Ⅱ(CN-Ⅱ)基因表达,以及长期传代后的变化规律,探索CN-Ⅱ与Ara-C疗效及耐药机制的相关性。方法采用RT-PCR方法测定HL-60和HL-60/R的细胞内CN-ⅡmRNA表达,并通过体外培养传代,检测长期传代(5、10、20、30、40、50代)后CN-ⅡmRNA表达变化。结果培养初期结果显示,HL-60和HL-60/R的CN-ⅡmRNA表达水平分别为0.23±0.04和0.47±0.05,HL-60/R的CN-ⅡmRNA表达水平明显高于HL-60(P<0.05)。经长期培养传代至10、20、30、40和50代,各传代检测节点的HL-60和HL-60/R细胞内CN-ⅡmRNA表达水平均无明显变化(P均>0.05)。上述各传代检测节点,HL-60/R的CN-ⅡmRNA表达水平均明显高于HL-60(P均<0.05)。结论 HL-60、HL-60/R细胞中CN-ⅡmRNA表达差异有统计学意义。经长期传代后,上述差异持续存在,提示CN-Ⅱ表达与Ara-C的疗效相关,但在长期传代后,变化并不明显,可能与Ara-C耐药现象的发生无关。

Abstract:

Objective To investigate the association of Cytosolic 5'-lucleotidase H (CN-H ) geneexpressions with drug resistant in long-term subcultured human leukemia HL-60 cells. Methods RealTime PCR was used to determinate the CN—n mRNA expressions in initial culture of HL-60 snd drug resisitant HL-60R cells and in their different passages (5，10，20， 30，40 and 50) . Results In the initial culture， the CN—n mRNA expression levels in HL-60 and HL-60/R cells were 0. 23 ±0. 04 and 0.47 ±0.05 (P <0. 05) . In the long-term subcultured passages (10，20，30，40，50)， the CN-H mRNA expression levels exhibited no change in both HL-60 and HL-60/R cells (P >0.05) . The CN-H mRNA expression levels in HL-60/R cell passages (10，20，30，40，50) were significantly higher than those in HL-60 cell passages (P <0.05) . Conclusion CN-H mRNA expression is higher in HL-60/Rcells than that in HL-60 cells, and the difference continuously exists after long-term cultured, which suggests that the CN-H expression is closely related to the development of drug resistance after long term use of Ara-C.