缺氧-复氧增强血小板介导的肿瘤细胞-内皮细胞黏附  

Hypoxia-reoxygenation exacerbated tumor cell adherence to endothelial cell mediated by platelet

作　　者：张娜[1] 刘虹麟[2] 蔡寒青[3] 彭亮[2] 李成辉[1] 叶丽亚[2] 许世清[2] 杨治华[1] 娄晋宁[2] 张文健[2] 

机构地区：[1]北京协和医学院研究生院,肿瘤研究所,北京100730 [2]中日友好医院临床医学研究所麻醉科,北京100029 [3]吉林大学第二医院内分泌科,吉林长春130041

出　　处：《中国癌症杂志》2011年第1期22-29,共8页China Oncology

基　　金：国家重点基础研究发展计划(No:2009CB521804);北京自然科学基金(No:7092093)

摘　　要：背景与目的:血小板能够与进入血液循环的肿瘤细胞黏附而促进其转移,但其机制是否涉及血小板作为介导物直接促进肿瘤细胞和内皮细胞黏附还不清楚,而且瘤栓在微血管局部造成的缺氧环境是否也具有促进作用尚不明确。本研究分析血小板对肿瘤细胞-内皮细胞黏附的影响及其相关黏附分子,以及缺氧-复氧在其中的作用。方法:将血小板和肿瘤细胞用荧光染料标记,通过荧光显微镜、连续光谱荧光仪以及激光共聚焦显微镜检测细胞的黏附情况;通过抗体阻断实验分析相关的细胞黏附分子,并在肠系膜血管上观察体内血小板促进肿瘤细胞在血管壁的黏附情况。结果:血小板与不同肿瘤细胞黏附能力不同,但都被缺氧-复氧所增强,并且不同肿瘤细胞与血小板的黏附被不同的抗血小板黏附分子的抗体阻断。人肝窦内皮细胞系(liver sinusoidal endothelial cell,LSEC)在缺氧-复氧后也明显增强与血小板的黏附,并且可以被抗黏附分子GPⅠb、GPⅡb、GPⅢa、P-selectin和CD31的抗体阻断。LSEC与血小板预温育可明显增加肿瘤细胞在其上的黏附,并且可以被抗血小板黏附分子的抗体阻断。激光共聚焦显微镜显示,血小板位于肿瘤细胞和LSEC之间,介导了两者的黏附。动物实验证实,与血小板预温育的肿瘤细胞在肠系膜血管上的黏附增多,并可以被缺血-再灌注进一步增强。结论:血小板表面存在与血管内皮细胞和肿瘤细胞同时黏附的分子,血小板通过这些黏附分子介导肿瘤细胞-内皮细胞的黏附,这种黏附作用可以被缺氧-复氧增强。Background and purpose： Adherence to platelets facilitates tumor cell metastasis but it is not clear whether platelets act as a mediator between tumor cells and endothelial cells or whether the hypoxia condition caused by tumor embolism enhances this process. The purpose of this study was to investigate the effect of platelets and hypoxia-reoxygenation on tumor endothelial cell adhesion and the role of relevant adhesion molecules. Methods： Platelets and tumor cells were labeled with fluorescent dye, then the adhesion were detected using a fluorescence microscope and fluorescence plate reader or laser scanning confocal microscope. The relevant adhesion molecules were analyzed by an antibody blockage experiment. The tumor cells adhesion to vessels in vivo were analyzed on mouse mesenteric vessels. Results： Hypoxia-reoxygenation promoted the adhesion of platelets to 4 different types of tumor cells. This could have been blocked by the antibodies against adhesion molecules on the platelets surface. Platelets adherence to human liver sinusoidal endothelial cells （LSEC） were also enhanced by hypoxia-reoxygenation and could be blocked by anti-GPIb, anti-GPIIb, anti-GPIIIa, anti-P-selectin and anti-CD31. Platelets increased the adhesion of tumor cells to LSEC, which could been blocked by antibodies against the adhesion molecules on the platelets. Confocal microscopy showed platelets present between tumor cells and LSEC. Animal experimentation indicated that tumor cell adherence to vessels was increased by platelets and further significantly enhanced when in a ischemia-reperfusion condition. Conclusion： Platelets could express that adhesion molecules adhered to both endothelial cells and tumor cells, and could also mediate the adhesion of tumor-endothelial cells. This adhesion could be enhanced through hypoxia- reoxygenation as well..

关 键 词：血小板 肝窦内皮细胞 肿瘤细胞 黏附 缺血-再灌注 

分 类 号：R730.23[医药卫生—肿瘤]