CIK逆转K562/ADR细胞多药耐药作用及其机制探讨

Reversion of multidrug resistance by CIK in K562/ADR cells and its mechanism exploration

目的 研究细胞因子诱导的杀伤细胞（CIK）体外逆转阿霉素（ADR）耐药细胞株K562/ADR细胞多药耐药（MDR）的作用,并探讨其机制.方法 健康人外周血单个核细胞经细胞因子体外诱导获得CIK,检测其表型和培养上清液细胞因子含量.实验组为CIK作用于K562/ADR细胞48 h后加入ADR;对照1组为CIK作用于K562/ADR细胞48 h,对照2组为ADR作用K562/ADR细胞48 h.采用MTT法检测各组细胞杀伤活性,用流式细胞术检测细胞膜P-糖蛋白（P-gp）含量、细胞内ADR浓度等.结果 实验组对K562/ADR细胞杀伤活性高于对照1组（P＜0.05）;且随效靶比增大,杀伤活性增大（P＜0.05）;随所加入的ADR浓度增大,杀伤活性无明显变化（P＞0.05）.实验组和对照1组的P-gp含量均下降（P＞0.05）.实验组细胞内ADR浓度高于仅经ADR作用的对照组2组（P＜0.05）,但细胞内ADR浓度与加入的ADR浓度无明显关系（P＞0.05）.结论 通过CIK与ADR对K562/ADR细胞的先后作用,降低了其细胞内P-gp的表达,提高了K562/ADR细胞内ADR浓度,增强了ADR对耐药细胞的杀伤活性.为应用生物活性细胞逆转MDR提供理论依据.

Objective To investigate the effects and mechanism of cytokine-induced killer（CIK） cells in reversing multidrug resistance（MDR） and increasing intracellular concentration of adriamycin（ADR）in the K562/ADR cells. Methods Peripheral mononuclear cells （MNCs） were isolated from healthy donors and cultured with combined cytokines to generate CIK. The changes of cell phenotype and cytokines secretion of CIK were determined. K562/ADR cells were divided into three groups： ADR in combination CIK （group Ⅰ ）, CIK alone （group Ⅱ ） and ADR alone （groupⅢ） . The viability and proliferation of K562/ADR cells were assayed by MTT assay, the intracellular concentration of ADR and the expression of P-glycoproteins （P-gp） in K562/ADR cells by FCM. Results The cytotoxicity of ADR in group Ⅰ was higher than that in group Ⅱ （ P 〈0.05 ）. The cytotoxicity was increased with the E/T ratio increasing（ P 〈0.05 ） but had no relation with the concentration of ADR in group Ⅰ （P〉0.05）. The expression of P-gp was declined in group Ⅰ and group Ⅱ （P 〉0.05 ）. The intracellular concentration of ADR in group Ⅰ was higher than that in group Ⅱ （ P 〈 0.05 ）, and had no relation with the ADR concentration （ P 〉 0.05 ）. Conclusion Pre-treatment with CIK can increase the cytotoxicity and the intracellular concentration of ADR and decrease the expression of P-gp in K562/ADR cells in the ADR and CIK combination group. Acute leukemia patients would be most likely to benefit from the combination of chemotherapy and CIK therapy.