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目的:通过建立鼻咽癌顺铂耐药细胞系CNE2/DDP,研究CXCR4在耐药机制中的作用。方法:采用药物浓度逐步递增的方法建立顺铂耐药细胞系CNE2/DDP,利用MTT实验、RNA干扰技术、microRNA过表达技术、荧光定量PCR和蛋白免疫印迹等分析CXCR4在CNE2/DDP顺铂耐药机制中的作用,并同时对其下游的目的基因进行研究。结果:1鼻咽癌顺铂耐药细胞系CNE2/DDP中CXCR4基因的表达水平升高,当降低了CXCR4siRNA表达水平后,CNE2/DDP的顺铂耐药能力减弱;2CNE2/DDP中的let-7a表达水平降低,而bcl-2的表达水平升高。当利用let-7amimics过表达let-7a后,bcl-2的表达水平下降,CNE2/DDP耐药能力减弱;3CXCR4siRNA降低CXCR4的表达水平后,let-7a的表达水平上升。结论:首次发现CNE2/DDP的耐药能力较CNE2明显增强,IC50值增加了至少5倍,并且通过调控let-7a的表达,进而影响凋亡抑制基因bcl-2的表达来实现其对CNE2顺铂耐药能力的调节。
OBJECTIVE: To investigate the role of CXCR4 in the mechanism of drug resistance by establishing a cell line CNE2 / DDP resistant to cisplatin of nasopharyngeal carcinoma. Methods: Cisplatin-resistant cell line CNE2 / DDP was established by gradually increasing the drug concentration. MTT assay, RNA interference technique, microRNA overexpression, fluorescent quantitative PCR and Western blotting were used to detect the expression of CXCR4 in CNE2 / DDP cisplatin- Drug mechanism in the role, and at the same time its downstream gene of interest to study. The expression of CXCR4 gene in CNE2 / DDP cell line was increased, and the cisplatin resistance of CNE2 / DDP group was weakened when the CXCR4siRNA expression was decreased. The let-|Ä- 7a expression decreased, while the expression of bcl-2 increased. After let-7a overexpression by let-7amimics, the expression of bcl-2 decreased and the drug resistance of CNE2 / DDP decreased. The expression of let-7a increased after 3CXCR4 siRNA decreased CXCR4 expression. CONCLUSION: The drug resistance of CNE2 / DDP was significantly enhanced compared with that of CNE2 for the first time, and the IC50 increased at least 5-fold. CNE2 / DDP was found to be resistant to CNE2 by regulating the expression of let-7a and then the expression of bcl- Platinum resistance to drug regulation.