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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 effect of cetuximab on the expression of natural killer group 2 member dligands (NKG2DLs) and the secretion of IFN-γ by NK cells in CNE2 / DDP cells. Methods: Flow cytometry was used to detect the expression of EGFR on CNE2 / DDP cells (ABCG2lowCNE2 / DDP cells and ABCG2highCNE2 / DDP cells) with high or low expression of ABCG2 (ATP-binding cassettesuperfamily G member 2) The expression of NKG2DLs on the surface of two CNE2 / DDP cells before and after cetuximab treatment. The two kinds of CNE2 / DDP cells before and after cetuximab were co-cultured with NK cells respectively. The secretion of IFN-γ in the supernatant was detected by ELISA and the killing of NK cells to different groups of CNE2 / DDP target cells by LDH release assay. Results: The expression rates of EGFR on ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP cells were (43.60 ± 2.01)% and (47.20 ± 2.07)%, respectively. Cetuximab up-regulated the expression of MICA, MICB, ULBP1 and ULBP2 on the surface of ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP cells, but down-regulated the expression of ULBP3 on ABCG2highCNE2 / DDP cell surface. Cetuximab treatment of two CNE2 / DDP cells, NK cell co-culture system with IFN-γ secretion was significantly increased (P <0.01); cetuximab enhanced two CNE2 / DDP cells NK Cell killing sensitivity (P <0.01). CONCLUSION: Cetuximab can up-regulate the expression of NKG2DLs in CNE2 / DDP cells and indirectly stimulate NK cells to secrete IFN-γ, which has a dual immunomodulatory effect.