细胞因子诱导杀伤细胞联合阿霉素逆转人红白血病耐药细胞株K562／ADR的研究  

Study of cytokine induced killer in combination with adriamycin reversing human rythroleukemia resistant cell line K562/ADR

作　　者：蒋东霞[1] 胡杰英[1] 杨晓博[1] 徐虹[1] 何琳[1] 买玲[1] 

机构地区：[1]河南省肿瘤研究所医学生物室,郑州450032

出　　处：《中国综合临床》2008年第3期225-227,共3页Clinical Medicine of China

基　　金：河南省科技厅专项基金资助项目（341700900）

摘　　要：目的探讨细胞因子诱导杀伤（CIK）细胞与阿霉素对肿瘤多药耐药逆转作用的影响。方法体外诱导、扩增人外周血单个核细胞（PBMC）作为效应细胞，联合不同浓度的阿霉素与其耐药靶细胞K562／ADR进行共培养，经四甲基偶氮唑盐（MTT）比色法计算半数抑制浓度（IC50）、耐药倍数和逆转倍数；流式细胞仪检测细胞内阿霉素相对含量和P糖蛋白（PgP）表达水平；RT—PCR检测mdr-1基因mRNA表达。结果阿霉素对K562和K562／ADR的IC50分别为（0．68±0．04）μmol／L和（66．23±4．38）μmol／L；K562／ADR对阿霉素的耐药倍数为97．43倍；经CIK细胞联合阿霉素处理48h（效靶比1：5、ADR1．1μmol／L）后，IC50为（32．15±0．79）μmol/L，联合逆转倍数为2．06倍。细胞内的阿霉素相对含量增加的同时PgP表达降低。mdr-1基因mRNA相对表达值也呈下调趋势。结论CIK细胞联合阿霉素可增加化疗药物阿霉素对耐药靶细胞K562／ADR的敏感性，提高耐药靶细胞内的阿霉素浓度，有效下调mdr-1基因及PgP的表达，使CIK联合阿霉素对多药耐药的逆转效果得以显现。Objective To explore the influence on the reversal role of cytokine induced killer （ CIK ） in combination with adriamycin （ ADR ）. Methods After separating mononuclear ceils from peripheral blood, effector ceils （ CIK） were induced and expanded in vitro. Combining with adriamycin-resistant target cell K562/ADR and adriamyein （ ADR ） in different concentrations, the 50 % inhibitory concentration （ IC50 ） , multiples of drug resistance and reverse were calealated according to the MTT assay; the concentration of intracellular ADR and expression level of P-gp were detected by flow cytometry; the expression level of mdr-1 mRNA was detected by semi-quantitative reverse transcriptase ploymerase chain reaction （PT-PCR）. Results IC50 of ADR in K562 and K562/ADR was （ 0.68 ± 0.04 ）μmol/L and （ 66.23 ± 4.38 ） μmol/L, the resistant multiple of K562/ADR to ADR was 97.43 ; With treatment of CIK in combination with ADR which lasted for 48 hours （ E/T = 1/5 ,ADR 1.1 μmol / L） ,the 50 % inhibitory concentration （IC50 ） was （32.15 ± 0.79）μmol/L,the joint reverse multiplier was 2.06. The concentration of intracellular ADR increased 1.83 times ; the expression of P-gp decreased 36.7% ; the expression of mdr-1 mRNA also showed a downward trend. Conclusion With CIK in combination with ADR, the sensitivity of ADR to K562/ADR could be increased, it also improves the concentration of intracellular ADR in K562/ADR and the expression level of mdr-1 mRNA and P-gp is downregulated,so that the reverse effect of CIK combined with ADR to muhidrug resistance is emerged.

关 键 词：细胞因子 杀伤细胞 阿霉素 多药耐药 K562/阿霉素细胞株 

分 类 号：R737.9[医药卫生—肿瘤]