文档介绍：PDTC逆转K562/AO2细胞多药耐药性机制研究
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【摘要】目的研究核因子κB(NF-κB)抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)逆转K562/AO2细胞耐药效应及其机制。方法采用MTT比色法分别检测K562/AO2细胞的耐药性、PDTC对K562/AO2细胞增殖的影响以及非细胞毒剂量PDTC、维拉帕米(Ver)预作用后K562/AO2细胞药物敏感性的变化,采用免疫细胞组织化学法、RT-PCR检测K562、K562/AO2细胞NF-κB、mdr-1 mRNA表达水平以及PDTC作用一定时间对其影响。结果①K562/AO2细胞对阿霉素(ADM)的耐药性是K562细胞的59倍;非细胞毒剂量PDTC预作用后,ADM对K562/AO2细胞的半数抑制浓度(IC50)显著降低,%,%();②K562/AO2细胞NF-κB表达水平高于K562细胞();PDTC可有效抑制K562/AO2细胞NF-κB表达,伴随mdr-1 mRNA表达减少,呈时间依赖性。结论抑制NF-κB异常活化可部分逆转K562/AO2细胞耐药性,其机制与mdr-1 mRNA转录表达减少有关。
【关键词】吡咯烷二硫代氨基甲酸盐;核因子
κB;K562/AO2细胞;多药耐药;mdr-1基因
Abstract: Objective To investigate the effect of pyrrolidine dithioearbama (PDTC), an inhibitor of nuclear factor κB (NF-κB) activity on reversing the chemoresistance of K562/AO2 cells and its mechanism. Methods Human erythroleukemic cell line K562 and its adriamycin-resistant counterpart K562/AO2 cells were used in the study. MTT assay was employed to detect the multiple drug resistance of K562/AO2 cells, the effect of PDTC on cell proliferation and the variations in drug sensitivity of K562/AO2 cells pretreated with PDTC/Verapamil (Ver) in noncytotoxic dose. Immunocytohistochemical method and RT-PCR were used to detect the relative expression of NF-κB and mdr-1 mRNA in K562 and K562/AO2 cells, respectively, and the effect of PDTC treatment at different time points. Results ①The drug resistance of K562/AO2 cells to ADM was 59 times as much as that of K562 cells. When pretreated with PDTC in noncytotoxic dose, the IC50 of ADM in K562/AO2 cells had a marked decrease, with a relative reverse efficiency of %, while that of classic modifying agents Ver was % (); ②The NF-κB expression level of K562/AO2 cells was signifi
cantly higher than that of K562 cells (); DTPC effectively inhibited the NF-κB expression of K562/AO2 cells, panied by the down-regulated expression of mdr-1 mRNA, in a time dependent manner. Conclusion With its inhibi