CXCR4介导的鼻咽癌细胞系CNE2对顺铂耐药性的研究  

Investigation of CXCR4 mediated chemoresistance in nasopharyngeal carcinoma cell line CNE2

作　　者：宋现运 尚小领[2] 张玉妥[1] 林彦涛[2] 徐国刚[2] 薛刚[2] 张青俊[2] 邹芳[1] 

机构地区：[1]河北北方学院病原微生物与免疫学研究所,河北张家口075000 [2]河北北方学院附属第一医院耳鼻咽喉头颈外科

出　　处：《临床耳鼻咽喉头颈外科杂志》2016年第11期887-891,896,共6页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

摘　　要：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：Since nasopharyngeal carcinoma is easy to develop resistance during cisplatin-based chemotherapy,CXCR4 expression levels were elevated in mang tumors,and the factor to do with tumor metastasis and chemotherapy drug resistance,and so on has a very important link.We established cisplatin-resistant nasopharyngeal carcinoma cell line,named as CNE2/DDP,and investigated the function of CXCR4 in molecular mechanism behind this resistance.Method：CNE2/DDP was firstly build up by increasing concentration of cisplatin.And then afterwards,MTT assay,RNA interference techniques,microRNA overexpresion techniques,quantative PCR and western blotting were applied to analyze the function of CXCR4 and its downstream effectors.Result：1the expression of CXCR4 was increased in CNE2/DDP and downregulation of CXCR4 with CXCR4siRNA was able to decrease the resistance of CNE/DDP to cisplatin;2the expression of let-7awas decrease in CNE2/DDP,while the expression of bcl-2was increased.Upregulation of let-7avia transfection of let-7amimics could downregulate the expression of bcl-2and damage the resistance of CNE2/DDP to cisplation;3downregulation of CXCR4 through CXCR4siRNA transfection was capable of improving the expression of let-7a.Conclution：We were the first to found that CXCR4 was related to chemoresistance of CNE2/DPP to cisplatin.Meanwhile,we confirmed that CXCR4 affected the expression of bcl-2through regulating the expression of let-7ato modulate the chemoresistance of CNE2/DPP to cisplatin.

关 键 词：鼻咽肿瘤 顺铂耐药性 CXCR4 let-7a bcl-2 

分 类 号：R739.63[医药卫生—肿瘤]