西妥昔单抗对耐药鼻咽癌CNE2/DDP细胞和NK细胞的双重免疫调节作用  

Dual immunological regulation effects of cetuximab on multidrug resistant nasopharyngeal carcinoma CNE2/DDP cells and NK cells

作　　者：黄宇贤[1] 王杨[2] 李玉华[1] 陈锦章[3] 钱敏[3] 吴秉毅[1] 孙彩霞[1] 邓兰[1] 郭坤元[1] 

机构地区：[1]南方医科大学珠江医院血液科,广东广州510282 [2]广东省人民医院肺癌研究所,广东广州510080 [3]南方医科大学南方医院肿瘤中心,广东广州510515

出　　处：《中国肿瘤生物治疗杂志》2010年第3期268-273,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.30973454);广东省自然科学基金重点项目(No.9251051501000007)~~

摘　　要：目的:探讨西妥昔单抗(cetuximab)对耐药鼻咽癌CNE2/DDP细胞NKG2D配体(natural killer group 2 member Dligands,NKG2DLs)的表达及NK细胞分泌IFN-γ的影响。方法:流式细胞术检测高、低表达ABCG2(ATP-binding cassettesuperfamily G member 2)的耐药鼻咽癌CNE2/DDP细胞(简称ABCG2lowCNE2/DDP细胞和ABCG2highCNE2/DDP细胞)表面EGFR的表达水平,以及西妥昔单抗处理前后两种CNE2/DDP细胞表面NKG2DLs的表达水平。西妥昔单抗处理前后的两种CNE2/DDP细胞分别与NK细胞共培养,ELISA检测上清中IFN-γ的分泌水平,LDH释放法检测NK细胞对不同组CNE2/DDP靶细胞的杀伤。结果:ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP细胞表面EGFR的表达率分别为(43.60±2.01)%和(47.20±2.07)%。西妥昔单抗上调ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP细胞表面MICA、MICB、ULBP1和ULBP2的表达,但下调ABCG2highCNE2/DDP细胞表面ULBP3的表达。西妥昔单抗处理两种CNE2/DDP细胞后,与NK细胞的共培养体系中IFN-γ的分泌水平明显上调(P<0.01);西妥昔单抗增强两种CNE2/DDP细胞对NK细胞杀伤的敏感性(P<0.01)。结论:西妥昔单抗可上调耐药鼻咽癌CNE2/DDP细胞NKG2DLs的表达,间接刺激NK细胞分泌IFN-γ,具有双重免疫调节作用。Objective To investigate the effects of cetuximab on NKG2D ligands（NKG2DLs） expressions in multidrug resistant nasopharyngeal carcinoma CNE2/DDP cells and IFN-γ production in NK cells.Methods： EGFR expressions on CNE2/DDP cells with high and low ABCG2 expression（ABCG2^highCNE2/DDP cells and ABCG2^lowCNE2/DDP cells） and NKG2DLs expressions on ABCG2^high and ABCG2^lowCNE2/DDP cells before and after cetuximab treatment were detected by flow cytomertry.ABCG2^high and ABCG2^lowCNE2/DDP cells were co-cultured with NK cells before and after cetuximab treatment,and then IFN-γ levels in the supernatants of different groups were detected by ELISA.Cytotoxicity of NK cells against CNE2/DDP cells was measured by LDH releasing assay in different groups.Results： EGFR expressions in ABCG2^high and ABCG2^lowCNE2/DDP cells were（43.60±2.01）% and（47.20±2.07）%,respectively.The expressions of MICA,MICB,ULBP1,and ULBP2 on ABCG2hgh and ABCG2^lowCNE2/DDP cells were up-regulated by cetuximab stimulation,while ULBP3 expression on ABCG2^highCNE2/DDP cells was down-regulated by cetuximab stimulation.IFN-γ levels in co-culture systems were significantly increased after ABCG2^low and ABCG2^highCNE2/DDP cells were treated with cetuximab（P〈0.01）.Cetuximab enhanced cytotoxic sensitivities of ABCG2^high and ABCG2^lowCNE2/DDP cells in response to NK cells（P〈0.01）.Conclusion： Cetuximab exerts a dual immunological regulation by up-regulating NKG2DLs expressions on nasopharyngeal carcinoma CNE2/DDP cells and stimulating IFN-γ production by NK cells indirectly.

关 键 词：西妥昔单抗 鼻咽肿瘤 多药耐药 NKG2D配体 自然杀伤细胞 

分 类 号：R730.54[医药卫生—肿瘤] R730.51[医药卫生—临床医学]