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目的:初步探讨舒尼替尼诱导高、低表达ABCG2(ATP-binding cassette superfamily G member 2)分子的耐药鼻咽癌CNE2/DDP细胞(简称ABCG2high CNE2/DDP细胞、ABCG2low CNE2/DDP细胞)中NKG2D配体(natural killer group 2 member Dligands,NKG2DLs)表达的分子机制。方法:Caspase-8活化试剂盒和线粒体膜电位法分别检测NK细胞处理后ABCG2highCNE2/DDP细胞和ABCG2lowCNE2/DDP细胞caspase-8活化水平和线粒体膜电位。RT-PCR检测舒尼替尼处理前后两种CNE2/DDP细胞DNA损伤修复系统相关信号分子mRNA的表达。结果:CNE2/DDP细胞+NK细胞组中两种CNE2/DDP细胞caspase-8活性均明显增强;舒尼替尼处理后的ABCG2lowCNE2/DDP细胞+NK细胞组和ABCG2highCNE2/DDP细胞+NK细胞组中caspase-8的活性是处理前的2～2.5倍(P<0.01)。舒尼替尼预处理后,CNE2/DDP细胞和NK细胞共培养体系中两种CNE2/DDP细胞的线粒体膜电位分别为(76.58±2.32)%和(73.11±1.93)%,较舒尼替尼处理前明显降低(P<0.05)。舒尼替尼可上调两种CNE2/DDP细胞中ATR、CHK1和CHK2 mRNA的表达,并诱导P53和NF-κB mRNA的表达。结论:舒尼替尼可能通过激活DNA损伤修复系统相关信号分子和NF-κB的表达,诱导耐药鼻咽癌CNE2/DDP细胞NKG2DLs的表达,同时经由死亡受体信号通路和线粒体信号通路增强NK细胞诱导的肿瘤细胞凋亡。
OBJECTIVE: To investigate the effects of sunitinib on CNE2 / DDP cells (ABCG2high CNE2 / DDP cells, ABCG2low CNE2 / DDP cells) with high expression of ABCG2 Molecular mechanisms of NKG2D ligand expression (NKG2DLs). Methods: Caspase-8 activation kit and mitochondrial membrane potential were used to detect caspase-8 activation and mitochondrial membrane potential in ABCG2highCNE2 / DDP cells and ABCG2lowCNE2 / DDP cells treated with NK cells. RT-PCR was used to detect the mRNA expression of related signal molecules of DNA damage repair system in both CNE2 / DDP cells before and after sunitinib treatment. Results: The activities of caspase-8 in both CNE2 / DDP cells in CNE2 / DDP cells + NK cells group were significantly increased. In ABCG2lowCNE2 / DDP cells + NK cells group and ABCG2highCNE2 / DDP cells + NK cells group The activity of caspase-8 was 2 to 2.5 times before treatment (P <0.01). After pretreatment with sunitinib, the mitochondrial membrane potentials of CNE2 / DDP cells in CNE2 / DDP cells and NK cells co-culture system were (76.58 ± 2.32)% and (73.11 ± 1.93)%, respectively, Significantly decreased before treatment (P <0.05). Sunitinib increased the expression of ATR, CHK1 and CHK2 mRNA in both CNE2 / DDP cells and induced the expression of P53 and NF-κB mRNA. Conclusion: Sunitinib may induce the expression of NF-κB and system-related signaling molecules in DNA damage-induced NPC cells and increase the expression of NKG2DLs in CNE2 / DDP cells, which may be mediated by death receptor and mitochondrial signal pathway Cell-induced apoptosis of tumor cells.