CXCR4介导的鼻咽癌细胞系CNE2对顺铂耐药性的研究

宋现运; 尚小领; 张玉妥; 林彦涛; 徐国刚; 薛刚; 张青俊; 邹芳

doi:10.13201/j.issn.1001-1781.2016.11.012

CXCR4介导的鼻咽癌细胞系CNE2对顺铂耐药性的研究

- 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

详细信息

通讯作者: 尚小领,E-mail:xiaolingshang66@sina.com

中图分类号: R739.63

收稿日期: 2015-12-25

Investigation of CXCR4 mediated chemoresistance in nasopharyngeal carcinoma cell line CNE2

- 1. Institute of Pathogen Biology and Immunology of Hebei North University, Zhangjiakou, 075000, China;
- 2. Department of Otolaryngology Head and Neck Surgery, the First Attached Hospital of Hebei North University

More Information

Corresponding author: SHANG Xiaoling, E-mail: xiaolingshang66@sina.com

Received Date: 25 December 2015

摘要

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CXCR4 siRNA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-7a mimics过表达let-7a后，bcl-2的表达水平下降，CNE2/DDP耐药能力减弱；③CXCR4 siRNA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。
- 鼻咽肿瘤 /
- 顺铂耐药性 /
- CXCR4 /
- let-7a /
- bcl-2
Abstract: 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 CXCR4 siRNA was able to decrease the resistance of CNE/DDP to cisplatin; ②the expression of let-7a was decrease in CNE2/DDP, while the expression of bcl-2 was increased. Upregulation of let-7a via transfection of let-7a mimics could downregulate the expression of bcl-2 and damage the resistance of CNE2/DDP to cisplation;③downregulation of CXCR4 through CXCR4 siRNA 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-2 through regulating the expression of let-7a to modulate the chemoresistance of CNE2/DPP to cisplatin.
- nasopharyngeal neoplasms /
- chemoresistance to cisplatin /
- CXCR4 /
- let-7a /
- bcl-2

HTML全文

参考文献(12)

[1]	AL-SARRAF M,REDDY M S.Nasopharyngeal carcinoma[J].Curr Treat Options Oncol,2002,3:21-32.
[2]	BURGER J A,KIPPS T J.CXCR4:a key receptor in the crosstalk between tumor cells and their microenvironment[J].Blood,2006,107:1761-1767.
[3]	MURDOCH C.CXCR4:chemokine receptor extraordinaire[J].Immunol Rev,2000,177:175-84.
[4]	DUDA D G,KOZIN S V,KIRKPATRICK N D,et al.CXCL12(SDF1alpha)-CXCR4/CXCR7 pathway inhibition:an emerging sensitizer for anticancer therapies[J]?Clin Cancer Res,2011,17:2074-2080.
[5]	LI X H,QU J Q,YI H,et al.Integrated analysis of differential miRNA and mRNA expression profiles in human radioresistant andradiosensitive nasopharyngeal carcinoma cells[J].PLoS One,2014,9:e87767.
[6]	CHEN Y,JACAMO R,KONOPLEVA M,et al.CXCR4 downregulation of let-7adrives chemoresistance in acute myeloid leukemia[J].J Clin Invest,2013,123:2395-2407.
[7]	JIN Y,SHI Y X,CAI X Y,et al.Comparison of five cisplatin-based regimens frequently used as the firstline protocols in metastatic nasopharyngeal carcinoma[J].J Cancer Res Clin Oncol,2012,138:1717-1725.
[8]	WANG T,MI Y,PIAN L,et al.RNAi targeting CXCR4 inhibits proliferation and invasion of esophageal carcinoma cells[J].Diagn Pathol,2013,8:104.
[9]	BARTEL D P.MicroRNAs:genomics,biogenesis,mechanism,and function[J].Cell,2004,23,116:281-297.
[10]	LEE Y S,DUTTA A.MicroRNAs in cancer[J].Annu Rev Pathol,2009,4:199-227.
[11]	MELTZER P S.Cancer genomics:small RNAs with big impacts[J].Nature,2005,435:745-746.
[12]	TSANG W P,KWOK T T.Let-7a microRNA suppresses therapeutics-induced cancer cell death by targeting caspase-3[J].Apoptosis,2008,13:1215-1222.