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 member2,ABCG2)耐药鼻咽癌细胞CNE2/DDP(ABCG2highCNE2/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ABCG2highCNE2/DDP细胞对Allo-NK细胞的杀伤敏感性明显增强。
Objective ： To investigate the inducing effects of sunitinib malate on expression of NKG2D ligands in nasopharyngeal carcinoma cell ABCG2high CNE2/DDP. Methods： ABCG2highCNE2/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 ABCG2highCNE2/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 ABCG2high 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.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2008年第4期311-315,共5页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.30471663)~~
