厄洛替尼作用后表皮生长因子受体突变的人肺腺癌HCC827细胞对细胞因子诱导的杀伤细胞的杀伤敏感性  

Susceptibility of human lung adenocarcinoma cancer HCC827 cells with epidermal growth factor receptor mutation to cytokine-induced killer cells after treated with erlotinib

作　　者：张世魁[1,2] 梅家转[2] 刘桂举 冯睿婷 

机构地区：[1]新乡医学院,河南新乡453003 [2]郑州人民医院肿瘤内科,河南郑州450003

出　　处：《新乡医学院学报》2015年第4期306-309,313,共5页Journal of Xinxiang Medical University

基　　金：郑州市科技创新领军人才基金项目(编号:121PLJRC532)

摘　　要：目的观察表皮生长因子受体(EGFR)酪氨酸激酶抑制剂厄洛替尼及EGFR下游主要分子对人肺腺癌HCC827细胞表面NKG2D配体表达的影响,探讨厄洛替尼作用于肺腺癌HCC827细胞后,细胞因子诱导的杀伤(CIK)细胞对HCC827细胞杀伤活性的变化。方法 HCC827细胞分为空白对照组及厄洛替尼5、10μmol·L-1组,应用流式细胞仪检测细胞表面NKG2D配体主要组织相容性复合体I类链相关蛋白(MIC)A、B及UL16结合蛋白(ULBP)1、2、3的表达。HCC827细胞培养24 h后,分别以EGFR下游分子磷脂酰肌醇3-激酶抑制剂LY294002、丝裂原活化蛋白激酶抑制剂SB203580、信号传导及转录激活因子3抑制剂STAT21、蛋白激酶C抑制剂Rottlerin进行作用,应用流式细胞仪分析HCC827细胞表面NKG2D配体MICA、MICB、ULBP1、ULBP2、ULBP3的表达,并与空白对照组进行比较。乳酸脱氢酶释放法检测CIK细胞对空白对照组和厄洛替尼组HCC827细胞的杀伤活性。结果与空白对照组比较,厄洛替尼5、10μmol·L-1组HCC827细胞表面NKG2D配体MICB、ULBP1表达显著增高(P<0.05),MICA、ULBP2、ULBP3表达差异无统计学意义(P>0.05);厄洛替尼5μmol·L-1组与10μmol·L-1组比较,HCC827细胞表面NKG2D配体MICA、MICB、ULBP1、ULBP2、ULBP3表达差异均无统计学意义(P>0.05)。与空白对照组比较,SB203580组的HCC827细胞表面NKG2D配体MICA、MICB、ULBP1、ULBP2、ULBP3表达差异无统计学意义(P>0.05),STAT21组的HCC827细胞表面NKG2D配体MICA、MICB表达显著增高(P<0.05),LY294002组的HCC827细胞表面NKG2D配体MICA表达显著降低(P<0.05),Rottlerin组的HCC827细胞表面NKG2D配体ULBP1表达显著增高(P<0.05)。同一效靶比时,CIK细胞对厄洛替尼作用后组HCC827细胞的杀伤率均显著高于未经药物作用组(P<0.05)。效靶比20∶1时经NKG2D单抗封闭后的CIK细胞对未经药物作用组和10μmol·L-1厄洛替尼作用后组HCC827细胞的杀伤率比较差异无统计学意义(P>0.05)。结论厄洛替尼�Objective To observe the effect of epidermal growth factor receptor（ EGFR） tyrosine kinase inhibitor erlotinib and EGFR downstream regulators on the expression of natural killer group 2,member D receptor（ NKG2D） ligands on human lung adenocarcinoma cancer HCC827 cells and detect the cytotoxicity mediated by cytokine-induced killer（ CIK） cells against HCC827 cells after treated with erlotinib. Methods The HCC827 cells were divided into blank control group and erlotinib 5,10 μmol·L- 1group. The expression of NKG2 D ligands major histocompatibility complex class I chain-related protein（MIC） A,B and UL16 binding protein（ULBP） 1,2,3 on HCC827 cells was detected by flow cytometery. After cultured for24 h and treated with the inhibitors of phosphatidylinositol 3-kinase LY294002,the inhibitors of mitogen-activated protein kinases SB203580,the inhibitors of signal transducers and activators of transcription 3 STAT21,and the inhibitors of protein kinase C rottlerin respectively,the expression of NKG2 D ligands（ MICA,MICB,ULBP1,ULBP2,ULBP3） was assayed by flow cytometery again,then the result was compared with the blank control group. Cytotoxicities of CIK cells against HCC827 cells in the blank control group and erlotinib group was detected by lactate dehydrogenase releasing assay. Results Compared with the blank control group,the expression of MICB and ULBP1 increased more significantly（ P 0. 05）,while the expression of MICA,ULBP2 and ULBP3 had not changed significantly（ P 0. 05） on HCC827 cells in the erlotinib 5,10 μmol·L- 1group.There was no significant difference of the expression of NKG2 D ligands（ MICA,MICB,ULBP1,ULBP2,ULBP3） between the erlotinib 5 μmol·L- 1group and 10 μmol·L- 1group（ P 0. 05）. There was no significant difference in the expression of NKG2 D ligands（ MICA,MICB,ULBP1,ULBP2,ULBP3） between the SB203580 group and the blank control group（ P 0. 05）,the expression of MICA and MICB in the STAT21 group and ULBP1 in the rottlerin group was significantly

关 键 词：肺癌 表皮生长因子受体酪氨酸激酶抑制剂 细胞因子诱导的杀伤细胞 NKG2D配体 

分 类 号：R392.12[医药卫生—免疫学]