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目的:探讨不同分子靶向药物对高表达与低表达ATP结合转运蛋白G超家族成员2(ATP-binding cassette super-family Gmember 2,ABCG2)的人耐药鼻咽癌CNE2/DDP细胞(分别简写为ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP)表面NKG2D配体(natural killer group 2 member Dligands,NKG2DLs)表达的诱导作用及其对NK细胞杀伤敏感性的影响。方法:免疫磁珠法分选ABCG2highCNE2/DDP、ABCG2lowCNE2/DDP细胞及NK细胞。流式细胞术检测分选细胞的纯度和不同分子靶向药物(硼替佐米、索拉非尼、舒尼替尼)处理前后ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP细胞NKG2DLs的表达率。LDH释放法检测不同药物处理前后ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP细胞对NK细胞杀伤的敏感性。结果:ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP细胞表面ABCG2的表达率分别为(91.40±2.32)%和(1.70±0.24)%。分选后NK细胞中CD3-CD16+CD56+细胞的比例达90%以上。药物处理前,ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP细胞MICA、MICB、ULBP1、ULBP2和ULBP3呈弱表达;经不同分子靶向药物处理后,5种NKG2DLs的表达率均明显上升(P<0.01),以舒尼替尼处理后NKG2DLs的表达率升高最明显。随着NKG2DLs表达的上调,ABCG2highCNE2/DDP和ABCG2lowCNE2/DDP细胞对NK细胞杀伤的敏感性也随之升高。结论:不同分子靶向药物可诱导耐药鼻咽癌CNE2/DDP细胞NKG2DLs的表达,以舒尼替尼的诱导作用最强,且肿瘤细胞NKG2DLs的表达与其对NK细胞杀伤敏感性之间存在线性关系。
OBJECTIVE: To investigate the effect of different molecular targeted drugs on CNE2 / DDP cells with high expression and low expression of ATP-binding cassette super-family G member 2 (ABCG2) (NKG2DLs) expression on ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP) and its effect on NK cell killing sensitivity. Methods: ABCG2highCNE2 / DDP, ABCG2lowCNE2 / DDP cells and NK cells were sorted by immunomagnetic beads method. Flow cytometry was used to detect the purity of sorted cells and the expression rates of NKG2DLs in ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP cells before and after treatment with different molecular targeted drugs (bortezomib, sorafenib, sunitinib). LDH release assay before and after treatment with different drugs ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP cells NK cell killing sensitivity. RESULTS: The ABCG2 expression rates on ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP cells were (91.40 ± 2.32)% and (1.70 ± 0.24)%, respectively. After sorting, the percentage of CD3-CD16 + CD56 + cells in NK cells was more than 90%. Before drug treatment, the expression of MICA, MICB, ULBP1, ULBP2 and ULBP3 in ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP cells were weakly expressed. The expression rates of 5 NKG2DLs were significantly increased after treatment with different molecular targeted drugs (P <0.01) Sunitinib treatment of NKG2DLs increased the most obvious expression rate. With the up-regulation of NKG2DLs expression, the sensitivity of NK cells to ABCG2highCNE2 / DDP and ABCG2lowCNE2 / DDP cells increased. CONCLUSION: Different molecular targeted drugs can induce the expression of NKG2DLs in CNE2 / DDP cells, and Sunitinib has the strongest inducing effect. There is a linear relationship between the expression of NKG2DLs and NK cell cytotoxicity relationship.