CXCR4及其配体SDF-1在宫颈癌转移中的作用及其机制研究  

The Role and Mechanism of CXCR4 and Its Ligand SDF-1 in the Development of Cervical Cancer Metastasis

作　　者：沈晓燕[1] 王绍海[1] 梁铭霖[1] 汪宏波[1] 肖兰[1] 王泽华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院妇产科,湖北武汉430022

出　　处：《癌症》2008年第10期1044-1049,共6页Chinese Journal of Cancer

基　　金：湖北省卫生厅科研基金(No.JXIA07)~~

摘　　要：背景与目的:趋化因子CXCR4[chemokine(C-X-C)receptor 4]受体及其配体基质细胞衍生因子1(stromal cell derived factor-1,SDF-1)可能与某些恶性肿瘤的迁移、侵袭和转移有关。本研究拟探讨CXCR4受体及其配体SDF-1(CXCR4/SDF-1轴)通过激活丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)信号通路在宫颈癌转移中的作用及其机制。方法:采用激光共聚焦显微镜检测宫颈癌HeLa细胞内钙离子波动,Western blot法分析CXCR4/SDF-1对宫颈癌HeLa细胞中细胞外调节信号激酶(extracellular signal-regulated kinase,ERK)(包括ERK1和ERK2)磷酸化的影响;分别通过粘附实验、明胶酶谱法检测CXCR4/SDF-1对宫颈癌细胞粘附能力、基质金属蛋白酶(matrix metalloproteinase,MMP)分泌的影响。结果:SDF-1α与CXCR4相互结合后诱导了HeLa细胞胞内钙的迅速动员,其荧光强度(fluorescent intensity,FI)平均基础值与峰值差异有统计学意义(P<0.01);诱导了HeLa细胞ERK1/2快速磷酸化,作用后30min时磷酸化程度最强;增加了HeLa细胞粘附能力,不同浓度的SDF-1α与CXCR4结合后不同程度地增加了HeLa细胞粘附于纤维连接蛋白(fibronectin,FN)、层粘连蛋白(laminin,LN)的粘附细胞数,与对照组比较其差异均有统计学意义(P<0.05);加入ERK1/2信号通路抑制剂PD98059后,粘附于FN和LN的细胞数下降,与对照组相比差异有统计学意义(P<0.05);促进宫颈癌HeLa细胞分泌活性的MMP-2,这一作用随SDF-1α浓度升高而增强,SDF-1α在800ng/mL浓度处分泌达高峰,以后开始下降。结论:CXCR4/SDF-1通过激活MAPK通路调控宫颈癌细胞的粘附能力,促进活性MMP-2分泌,进而参与宫颈癌侵袭和转移。BACKGROUND ＆ OBJECTIVE： The chemokine receptor CXCR4 and its sole ligand stromal cell-derived factor-1 （SDF-1） not only actively participate in inflammation, hematopoiesis, infection of HIV, but also play a pivotal role in migration, invasion and metastasis of some malignant tumors. This study was to investigate the role of CXCR4/SDF-1 axis in mediating metastasis in cervical cancer cells through activating the mitogenactivated protein kinase （MAPK） pathway and its possible mechanism. METHODS. Intracellular calcium mobilization was observed under laser scanning confocal fluorescence microscopy. The phospharylation of extracellular singal-regulated kinase （ERK） 1/2 in HeLa cells after binding of SDF-la to CXCR4 was measured by Western blot. Adhesion of CXCR4/SDF-1 to cervical cancer cells and secretion of matrix metalloproteinase （MMP） were detected by adhesion assay and gelatin zymography, respectively. RESULTS. After SDF-la was bound to CXCR4, a rapid and robust mobilization of intracellular calcium in Hela cells was initiated. The difference between the average baseline fluorescence intensity （FI） and the peak FI was significant （P 〈 0.01 ）. ERK-1/2 was rapidly phosphorylated in Hela cells after its exposure to SDF-la, and the strongest phosphorylation occurred at 30 min. The adhesion ability of Hela cells to fibronectin （FN） and laminin （LN） was increased after SDF-la treatment （P〈0.05 and P〈0.01, respectively）, while pretreatment of Hela cells with an ERK-1/2 inhibitor, PD98059, decreased adhesion of Hela cells to extracellular matrix （ECM） with the presence of SDF-la （P〈0.05）. Increased amounts of active MMP-2 were secreted in response to increased SDF-la concentrations. When the concentration of SDF-la was 800ng/mL, the secretion of active MMP-2 reached the peak and started to decrease afterwards. CONCLUSION： CXCR4/SDF-1 participate in tumor invasiveness and metastasis in cervical cancer through regulating the adhesion ability by activating the MAPK

关 键 词：CXCR4 SDF-1 宫颈肿瘤 HELA细胞 转移 

分 类 号：R737.33[医药卫生—肿瘤]