ET-1通过上调CXCR4表达促进鼻咽癌低转移细胞株6-10B的迁徙  

Endothelin-1 Promotes 6-10B Nasopharyngeal Carcinoma Cell Migration by Inducing the Expression of Chemokine Receptor CXCR4

作　　者：麦海强[1] 徐理华[2] 刘怀[1] 莫浩元[1] 黄培钰[1] 唐林泉[1] 陈秋燕[1] 

机构地区：[1]华南肿瘤学国家重点实验室//中山大学肿瘤防治中心鼻咽癌科,广东广州510060 [2]华南肿瘤学国家重点实验室//中山大学肿瘤防治中心中心实验室,广东广州510060

出　　处：《中山大学学报（医学科学版）》2011年第5期616-622,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金项目(3060075581072226);"863计划"子课题(2006AA02A404);广东省科委项目(2007B060401064);广东省医学科研基金(B2006063);广州市科委项目(2007Z1-E4022);中央高校基本科研业务费专项资金(2009)

摘　　要：【目的】探讨内皮素-1(ET-1)上调CXCR4表达促进鼻咽癌低转移细胞株6-10B迁徙的机制。【方法】加入不同浓度的ET-1以及同时加入SDF-1α处理6-10B细胞,趋化实验检测ET-1对6-10B细胞迁徙能力的影响。分别采用Real-time PCR以及Western Blot方法检测ET-1上调后CXCR4在基因以及蛋白水平的变化。Western Blot方法检测用特异性的ETAR抑制剂或者PI3K/AKT/mTOR或MARK1/ERK1/2抑制剂预处理6-10细胞后,ET-1对CXCR4表达水平的变化;并探讨ET-1处理后,PI3K/AKT/mTOR以及MARK1/ERK1/2信号通路中蛋白磷酸化水平的改变。【结果】趋化实验显示6-10B细胞丧失对SDF-1α的趋化能力,但不同浓度的ET-1刺激后6-10B细胞对SDF-1α的迁徙能力明显增强(P<0.05),10 nmol/L ET-1的作用最明显。Real-time PCR以及Western Blot结果显示,不同浓度的ET-1处理6-10B细胞后,ET-1在基因以及蛋白的水平可以呈剂量依赖性及时间依赖性地上调CXCR4的表达。用特异性的内皮素受体A(ETAR)抑制剂或者PI3K/AKT/mTOR或MARK1/ERK1/2通路的抑制剂,能够明显抑制ET-1对CXCR4表达的上调。Western blot检测显示ET-1能激活PI3K/AKT/mTOR及MAPK1/ERK1/2信号通路。【结论】ET-1能上调6-10B细胞功能性CXCR4的mRNA和蛋白表达,并能明显增强6-10B细胞对SDF-1α的迁徙能力。CXCR4的上调主要是通过ETAR介导,激活PI3K/AKT/mTOR和MAPK1/ERK1/2信号通路。【Objective】 To evaluate the mechanism of Endothelin-1（ET-1） promoting 6-10B nasopharyngeal carcinoma cell migration by inducing the expression of CXCR4.【Methods】 Chemotaxis assays were used to evaluate the migration ability of 6-10B after 6-10B cells were treated with different concentration of ET-1 together with SDF-1α.Real-time PCR and Western Blot were used to evaluate mRNA and protein expression change of CXCR4 induced by ET-1.Western Blot was conducted to detect the expression change of CXCR4 induced by ET-1 when 6-10B cells were pre-treatment by Endothelin A receptor（ETAR） or PI3K/AKT/mTOR or MARK1/ERK1/2 inhibitor.Detect the protein phosphorylation expression change in the pathway of PI3K/AKT/mTOR or MARK1/ERK1/2 after treated with ET-1.【Results】 Chemotaxis assays showed that the migration of 6-10B cells had no response to SDF-1α.Different concentration of ET-1 together with SDF-1 promoted the migration ability of 6-10B（P 0.05）,especially when the ET-1 concentration was 10 nmol/Lol/L.Real-time PCR and Western Blot results showed that ET-1 induced CXCR4 mRNA and protein expression in 6-10B nasopharyngeal carcinoma cells in a time-and concentration-dependent fashion.ET-1-induced CXCR4 expression could be inhibited by ETAR or PI3K/AKT/mTOR or MARK1/ERK1/2 inhibitor.Western Blot results showed that ET-1 inducing CXCR4 expression could activate the PI3K/AKT/mTOR or MARK1/ERK1/2 signal pathways.【Conclusion】 ET-1 induced the mRNA and protein expression of CXCR4 and promoted the migration ability of 6-10B to SDF-1αvia mediated through ETAR and activated by PI3K/AKT/mTOR or MARK1/ERK1/2 signal pathways.

关 键 词：鼻咽癌 ET-1 CXCR4 迁徙 

分 类 号：R739[医药卫生—肿瘤]