顺铂与CIK细胞对A549细胞杀伤协同作用机制的初步研究  

Sequential effect of Cisplatin on killing effect of CIK cell to human lung adenocarcinoma A549 cell line

作　　者：张晓娟[1] 梅家转[1] 赵继智[1] 冯睿婷 刘桂举 

机构地区：[1]郑州人民医院肿瘤内科,河南郑州450003

出　　处：《中华肿瘤防治杂志》2013年第16期1229-1231,共3页Chinese Journal of Cancer Prevention and Treatment

基　　金：郑州市科技领军人才项目(121PLJRC532)

摘　　要：目的:探讨不同浓度顺铂作用人肺腺癌A549细胞24h后,NKG2D配体表达的改变及细胞因子诱导的杀伤细胞(CIK)杀伤活性的变化。方法:MTT法测定顺铂作用A549细胞24h的50%抑制浓度(IC50);流式细胞仪检测IC50、1/2IC50和1/4IC50浓度顺铂分别作用A549细胞24h后,A549细胞表面NKG2D配体表达的变化,配体包括MHC-I类链相关分子MICA和MICB以及人巨细胞病毒糖蛋白UL16结合蛋白ULBP1、ULBP2和ULBP3;乳酸脱氢酶释放法检测不同效靶比时,CIK细胞对IC50浓度的顺铂作用前及作用24h后A549细胞的杀伤活性。结果:不同浓度顺铂作用24h后,A549细胞表面MICA(F=12.490,P=0.002)、MICB(F=41.492,P<0.001)、ULBP2(F=375.773,P<0.001)和ULBP3(F=59.594,P<0.001)表达均显著升高;ULBP1表达降低,F=55.693,P<0.001。效靶比10∶1时,IC50浓度顺铂作用24h前、后的A549细胞的杀伤活性分别为(11.88±1.57)%和(17.64±1.44)%,F=21.770,P=0.010;20∶1时分别为(35.56±1.98)%和(46.39±3.51)%,F=21.653,P=0.010;30∶1时分别为(45.03±1.74)%和(73.81±1.62)%,F=439.578,P<0.001。结论:顺铂提高A549细胞NKG2D配体MICA、MICB、ULBP2和ULBP3的表达,增强A549细胞对CIK细胞杀伤的敏感性。OBJECTIVE：To analyse the effects of cisplatin on the expression of NKG2D ligands and the cytotoxicity of cytokine-induced killer （CIK） ceils against human lung adenocarcinoma cell line A549 after 24 h treated with different concentrations of cisplatin. METHODS .. The IC50 of cisplatin against A549 ceils were measured by MTT assay. Expression of NKG2D ligands （MICA, MICB,ULBP1, ULBP2, ULBP3） on surface of A549 cell before and after 24 h treated by cis- platin （ICso, 1/2 IC50,1/4 IC50 ） were assaied by flow cytometery. Cytotoxicities of CIK ceils against A549 ceils were ana- lyzed by LDH releasing assay at effector -to-target cell ratio （E ： T） of 10 - 1,20 ： 1,30 ： 1. RESULTS： Expressions of MICA,MICB,ULBP2, ULBP3 were increased in A549 cells treated with different concentrations of cisplatin for 24 h （F values were 12. 490,41. 492,375. 773,59. 594,all P〈0.05） ,while ULBP1 was decreased （F=55. 693,P〈0. 001）. Cytotoxicity of CIK cells against A549 cells were （11.88±1.57）%, （35.56± 1.98） %, （45.03±1.74）% at the E ：T ra- tios of 10 ： 1,20 ： 1,30 ： 1,respectively,which were significantly increased after treated with IC50 cisplatin （F= 21. 770, 21. 653,439. 578, all P〈0.05 ）, which were （ 17.64 ±1.44）%, （46.39 ± 3.51 ）% and （73.81 ± 1.62）%, respectively. CONCLUSION ： This study indicates that cisplatin enhances the susceptibility to CIK cell-mediated cytotoxicities by upreg- ulating NKG2D ligands in A549 cells.

关 键 词：肺癌 顺铂 细胞因子诱导的杀伤细胞 NKG2D配体 化疗 

分 类 号：R734.2[医药卫生—肿瘤]