HL60/VCR白血病细胞多药耐药机制的初步探讨

Study on mechanisms of multidrug resistance in HL60/VCR leukemia cell line

目的探讨白血病细胞产生耐药的机制,以及VEGF在白血病细胞产生耐药过程中的作用。方法采用反复暴露并逐渐提高诱导药物浓度的方法建立HL60/VCR多药耐药细胞系,MTT法检测细胞的耐药性及交叉耐药性;RT-PCR检测诱导前后mdr1、MRP、TopoⅡα、GSTπ和VEGF mRNA的表达情况。结果诱导后的HL60/VCR耐药细胞不仅对VCR高度耐药,对其他类化疗药物亦具有交叉耐药性;与HL60相比,HL60/VCR的mdr1、TopoⅡα和VEGF mRNA的表达明显增强,GSTπmRNA的表达无明显变化;HL60和HL60/VCR均不表达MRP mRNA。结论耐药基因mdr1和TopoⅡα以及VEGF共同参与HL60/VCR多药耐药的形成。

Objective To investigate the mechanisms underlying multi-drug resistance（MDR） of leukemia cells ,as well as the effect of VEGF in the MDR of leukemia cells. Methods To induce human promyeloid leukemia line （HL60） into multi-drug resistant cell line HL60/VCR, an intermittent vincristine was added step by step into the culture medium of the tumor cells and the concentration of vincristine was gradually increased. MTT assay was used to detect the sensitivity of these tumor cells to different sorts of chemotherapeutic agents. The mRNA expression of mdr1, MRP, TopoⅡα, GSTπ and VEGF was determined by reverse transcriptase polymerase chain reaction （RT-PCR）. Results Compared to sensitive HL60 cells,HL-60/VCR cells were more resistant to VCR and other different sorts of chemotherapeutic agents. The expression of mdr1,TopoⅡα and VEGF mRNA of HL60/VCR cells was significant higher than those of HL60, while no difference was found in the expression of GSTπmRNA. Both HL60 and HL60/VCR did not express MRPmRNA. Conclusion mdr1,TopoⅡα and VEGF play important role in the MDR of HL60/VCR.