摘 要 目的: 探讨苹果酸舒尼替尼(sunitinib malate, SU11248)对高表达三磷酸腺苷结合转运蛋白G超家族成员2(ATP-binding cassette superfamily G member 2,ABCG2)耐药鼻咽癌细胞CNE2/DDP(ABCG2high CNE2/DDP)表达NKG2D配体的诱导作用。方法: 利用免疫磁珠技术分离ABCG2highCNE2/DDP细胞及同种异体反应性自然杀伤细胞(allo-reactive natural kill cell, Allo-NK)，流式细胞技术检测分离后细胞的纯度及苹果酸舒尼替尼处理前后ABCG2highCNE2/DDP细胞NKG2D配体表达率，LDH释放测定法检测苹果酸舒尼替尼处理前后ABCG2highCNE2/DDP细胞对Allo-NK细胞的杀伤敏感性。结果: ABCG2highCNE2/DDP细胞分离后ABCG2的表达率为(91.40±2.32)%，Allo-NK细胞分选后CD3－CD16+CD56+细胞的纯度达90%以上。经苹果酸舒尼替尼处理后,ABCG2highCNE2/DDP细胞的NKG2D配体MICA、MICB、ULBP1、ULBP2、ULBP3的表达率由药物处理之前的(2.92±0.33)%、(4.27±0.33)%、(5.80±0.62)%、(11.10±3.15)%、(7.75±1.14)%分别上升到(89.12±4.56)%、(66.10±2.22)%、(67.56±4.19)%、(69.37±8.83)%、(63.28±3.31)%。在效靶比为10∶1、20∶1时，苹果酸舒尼替尼处理前后Allo-NK细胞对ABCG2highCNE2/DDP细胞的杀伤率分别为(15.32±13.86)%、(27.26±6.81) % 及(41.12±4.12)%、(57.25±2.37)%，处理后的杀伤率有明显的提高(F=15.58， P＝0.000)。结论: 苹果酸舒尼替尼通过诱导高表达NKG2D配体(MICA/B、ULBP1-3)，使ABCG2high CNE2/DDP细胞对Allo-NK细胞的杀伤敏感性明显增强。

Abstract Objective: To investigate the inducing effects of sunitinib malate on expression of NKG2D ligands in nasopharyngeal carcinoma cell ABCG2high CNE2/DDP. Methods:ABCG2highCNE2/DDP cells and AlloNK cells were isolated by magnetic activated cell sorting (MACS). Flow cytometry was used to evaluate the purity of isolated cells and the expression of NKG2Dligands on target cells before and after incubation with sunitinib malate. Then the cytotoxic sensitivity of treated and untreated ABCG2high CNE2/DDP cells to AlloNK cells were measured by LDH releasing assay. Results: The positive rate of ABCG2 in ABCG2highCNE2/DDP cells was (91.40±2.32)%. More than 90% of isolated AlloNK cells were proven to be CD3-CD16+CD56+ cells. The expression of MICA, MICB,ULBP1, ULBP2 and ULBP3 on ABCG2high CNE2/DDP cells incubated with sunitinib malate increased from (2.92±0.33)%, (427±0.33)%, (5.80±0.62)%, (11.10±3.15)%, and (7.75±1.14)% to (89.12±4.56)%, (66.10±222)%, (67.56±4.19)%, (69.37±8.83)%, and (63.28±3.31)%, respectively. At the E∶T ratios of 10∶1 and 20∶1, the cytotoxic sensitivities of ABCG2high CNE2/DDP cells to AlloNK cells increased from (15.32±13.86)% and (27.26±6.81)% to (41.12±4.12)% and (57.25±2.37)%, respectively, after treatment with sunitinib malate, with significantly difference found in the cytotoxic sensitivities of target cells in each group before and after sunitinib malate treatment (F=1558, P＝0.000). Conclusion: Sunitinib malate can upregulate expression of NKG2Dligands (MICA/B, ULBP13) in ABCG2high nasopharyngeal carcinoma cells, which results in higher cytotoxic sensitivity to AlloNK cells.

国家自然科学基金资助项目（No.30471663）.