【作者】 李蒲；

【导师】 汤爱萍；

【作者基本信息】 南昌大学 ， 内科学， 2007， 硕士

【摘要】 目的:探讨干扰素(IFN-α)和白细胞介素-2(IL-2 )对人类髓性白血病多药耐药细胞株HL-60/VCR的作用及作用机制,为IFN-α和IL-2在难治性急性白血病中的应用提供实验依据。方法:采用甲基四氮唑盐(MTT)法检测IFN-α和IL-2对多药耐药细胞株HL-60/VCR细胞生长的抑制作用;采用流式细胞术检测IFN-α、IL-2、IFN-α联合IL-2处理前后细胞中的P-gp表达水平的变化,进一步探讨其逆转耐药的机制。结果:1.HL-60细胞及其多药耐药株HL-60/VCR细胞对VCR药物的敏感性分别IC50=3.49ng/ml、IC50=926.12 ng /ml。其HL-60/VCR细胞对VCR的耐药倍数是HL-60细胞的265.57倍,耐药倍数较高,符合实验要求。2.IFN--α浓度在0-500U/ml之间时对HL-60/VCR细胞未见明显的抑制作用,当细胞因子浓度>500U/ml其抑制作用明显加强,并随剂量加大作用加强。3.IL-2浓度为0-1000U/ml范围内对HL-60/VCR细胞未见明显的抑制作用。4.IL-2、IFN-α及IL-2联合IFN-α对HL-60/VCR细胞均有一定的逆转耐药作用,逆转倍数分别为1.53、1.90、3.62倍,且VCR浓度在0.1-2.0范围内IFN-α联合IL-2的逆转作用较细胞因子单独使用时作用明显增强,具有显著性差异,P<0.05。5.HL-60/VCR细胞在IFN-α、IL-2及IFN-α联合IL-2作用前后的细胞表面P-gp表达水平分别为33.41±4.63,23.07±5.48,21.82±3.87,16.62±1.27,IFN-α组和IL-2+IFN-α组明显下降,P<0.05;细胞内总P-gp的表达水平分别为37.19±5.16、35.3±4.77、37.45±5.19、34.98±3.50,各组间无显著差异,P>0.05。结论:1.HL-60/VCR细胞为人类髓细胞性白血病多药耐药株,其对VCR有较高耐药性。2.IFN-α在终浓度均为0-500U/ml时对HL-60/VCR细胞的增殖无明显的抑制作用,但IFN-α终浓度>500U/ml时出现了明显的生长抑制作用,说明高浓度IFN -α对HL-60/VCR具有一定的杀伤作用。3.在0-1000U/ml终浓度范围内的IL-2对HL-60/VCR细胞无明显的杀伤作用。4.IFN-α和IL-2与VCR同共作用于HL-60/VCR细胞,其对VCR的敏感性增高,说明IFN-α和IL-2具有部分逆转白血病细胞MDR作用。5.IFN-α联合IL-2(终浓度均为500U/ml)作用HL-60/VCR细胞后,其逆转作用增强。6.IFN-α联合IL-2应用逆转MDR作用的机制一部分可能是通过影响细胞质和细胞膜之间的P-gp的交换,减少细胞膜上P-gp表达水平,提高药物在细胞内的聚集,增强药物对肿瘤细胞的敏感性从而发挥杀死肿瘤细胞的作用。更多还原

【Abstract】 Objectives: To investigate the effect of IFN-a plus IL-2 on multidrug resistance of myelogenous leukemia cell line HL-60/VCR and the underlying mechanisms.Methods: The inhibitory effect of IFN-a plus IL-2 on HL-60/VCR growth was detected by MTT . The expression levels of P-gp in HL-60/VCR cells treated in the presence or absence of IFN-a, IL-2, IFN-a plus IL-2 respectively were determined by Flow Cytometry ,To research machine of resistance retroconversion of leukemia cellResults: 1. IC50 of VCR to HL-60 and HL-60/VCR are 3.49ng/ml and 0.92μg/ml respectively. Multidrug resistance of HL-60/VCR is 265.57times than that of HL-60, which accords with the requirement of the study.2.IFN-a shows no significant inhibitory effect on HL-60/VCR at the concentration varies from 0U/ml to 500U/ml, but shows significant effect in the dose of more than 500U/ml. The inhibitory effect was in a dose dependent manner.3. IL-2 shows no significant inhibitory effect on HL-60/VCR at the concentration varies from 0U/ml to 1000U/ml.4.All the IFN-a, IL-2, IFN-a plus IL-2 can induce drug resistance retroconversion, and the retroconversion index are1.53,1.90,3.62times respectively. When the concentration of VCR varies from 0.1 to 2.0, the inhibitory effect of IFN-a plus IL-2 is significantly higher than that of the cytokine alone (P<0.05) ).5.The expression level of P-gp in HL-60/VCR cells treated in the presence or absence of IFN-a plus IL-2 was. 33.41±4.63,23.07±5.48,21.82±3.87,16.62±1.27 respectively,group IFN-a and IL-2+IFN-αare obvious decent, P<0.05; The total expression level of P-gp in cells37.19±5.16、35.3±4.77、37.45±5.19、34.98±3.50 respectively,each interclass no significant deviation P>0.05。Conclusions:1.HL-60/VCR, a myelogenous leukemia cell line, have multidrug resistance to VCR.2.IFN-αshows no significant inhibitory effect on HL-60/VCR proliferation at the concentration varies from 0U/ml to 500U/ml, but shows significant effect in the dose of more than 500U/ml. the result indicate that high dose of IFN-αable to impair HL-60/VCR proliferation.3.IL-2 can not impair HL-60/VCR proliferation at the concentration varies from 0U/ml to 1000U/ml.4.IFN-αand IL-2 can accentuate the effect of VCR on HL-60/VCR, which indicates IFN-αand IL-2 can induce multidrug resistance retroconversion.5.The inhibitory effect of IFN-αplus IL-2(500U/ml) is significantly higher than that of IFN-αor IL-2 alone.6.IFN-αplus IL-2 may induce multidrug resistance retroconversion and kill the leukemia via influencing exchange of P-gp between cytoplasm and cytomembrane of HL-60/VCR, attenuating the expression of P-gp on cytomembrane, promoting the aggregation of VCR in cytoplasm and enhancing the effect of medicine on HL-60/VCR..更多还原