吉非替尼上调NKG2D配体表达增强A549细胞对NK细胞杀伤的敏感性

摘要

Objective:To explore the effects of gefitinib on the expression of NKG2D ligands in human lung cancer A549 cells and the cytolytic activity of NK cells. Methods: MTT method was used to detect the anti-proliferative ratio of gefitinib on A549 cells. Expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) on A549 cells before and after treated with gefitinib, LY294002 (PD-K inhibitor ) , SB203580 (MAPK inhibitor) , STAT21 (STAT3 inhibitor) , Rottlerin (PKC inhibitor) were analysed respectively by flow cytometer. Cytotoxicities of NK cells against A549 cells before and after treated with gefitinib were measured by LDH releasing assay at effect-to-target cell ratios of 5- 1, 10: 1, 20: 1. Results: Expressions of MICB and ULBP1 were increased in A549 cells treated with gefitinib. Effect of MAPK inhibitors ( SB203580) and STAT3 inhibitors ( STAT21) on expressions of NKG2D ligands were not significant, while PD-K inhibitors (LY294002) downregulated expression of MICA. PKC inhibitors (Rottlerin ) increased expression of ULBP1. Cytotoxicity of NK cells against A549 cells treated by gefitinib was significantly enhanced (P <0. 05). Conclusion:This study indicates that gefitinib enhances the susceptibility to NK cell-mediated cytotoxicities by upregulating NKG2D ligands in A549 cells.%目的:研究表皮生长因子受体酪氨酸激酶抑制剂吉非替尼对人肺腺癌A549 细胞NKG2D配体表达及NK细胞杀伤活性的影响及其分子机制.方法:MTT法测定吉非替尼对A549细胞增殖抑制率,流式细胞仪检测吉非替尼、EGFR下游分子LY294002 (PI3-K抑制剂) 、SB203580(MAPK抑制剂) 、STAT21(STAT3抑制剂)、Rottlerin (PKC抑制剂)作用A549细胞24小时后A549细胞NKG2D配体的表达.乳酸脱氢酶释放法检测不同效靶比时,NK细胞对吉非替尼作用前、后A549细胞的杀伤活性.结果:吉非替尼上调A549细胞MICB、ULBP1表达,增强A549细胞对NK细胞杀伤的敏感性,EGFR下游分子MAPK、STAT3抑制剂不影响A549细胞NKG2D配体的表达,PI3-K抑制剂下调A549细胞MICA表达,PKC抑制剂上调ULBP1表达.结论:吉非替尼上调NKG2D配体表达增强A549细胞对NK细胞杀伤的敏感性.