CAJ | 학술논문

目的体外分析黏附分子αvβ3和αvβ5及其配体Del-1、L1在肿瘤细胞-内皮细胞黏附中的作用.方法应用逆转录聚合酶链反应(RT-PCR)和流式细胞分析比较正常肝窦内皮细胞(LSEC)和肝癌的血管内皮细胞T3A上细胞间黏附分子1(ICAM-1)、αvβ3和αvβ5的表达以及缺氧对其表达的调控.分别用RT-PCR和Western blot方法分析6种肿瘤细胞Del-1和L1的表达及缺氧对其表达的调控.使用连续光谱荧光测定仪定量肿瘤细胞在LSEC和T3A上的黏附,并分析抗不同黏附分子的抗体和siRNA对肿瘤细胞一内皮细胞黏附的阻断作用.结果 T3A细胞αvβ3和αvβ5的基础表达高于LSEC,而ICAM-1的基础表达低于LSEC.缺氧上调两种内皮细胞αvβ3和αvβ5的表达,而ICAM-1表达仅在LSEC中升高.在不同肿瘤细胞中,Del-1和L1的基础表达存在明显差异,并且在缺氧条件下受到明显不同的调节.Del-1和L1高表达的肿瘤细胞在T3A细胞的黏附明显高于LSEC,并且在缺氧条件下黏附明显增加,这种黏附可以被抗αvβ3、αvβ5的抗体或沉默β3和β5的小干扰RNA(siRNA)阻断.结论细胞黏附分子αvβ3、αvβ5及其配体在肿瘤细胞-肿瘤内皮细胞的黏附过程中起重要介导作用.
목적 체외분석점부분자αvβ3화αvβ5급기배체Del-1、L1재종류세포-내피세포점부중적작용.방법 응용역전록취합매련반응(RT-PCR)화류식세포분석비교정상간두내피세포(LSEC)화간암적혈관내피세포T3A상세포간점부분자1(ICAM-1)、αvβ3화αvβ5적표체이급결양대기표체적조공.분별용RT-PCR화Western blot방법분석6충종류세포Del-1화L1적표체급결양대기표체적조공.사용련속광보형광측정의정량종류세포재LSEC화T3A상적점부,병분석항불동점부분자적항체화siRNA대종류세포일내피세포점부적조단작용.결과 T3A세포αvβ3화αvβ5적기출표체고우LSEC,이ICAM-1적기출표체저우LSEC.결양상조량충내피세포αvβ3화αvβ5적표체,이ICAM-1표체부재LSEC중승고.재불동종류세포중,Del-1화L1적기출표체존재명현차이,병차재결양조건하수도명현불동적조절.Del-1화L1고표체적종류세포재T3A세포적점부명현고우LSEC,병차재결양조건하점부명현증가,저충점부가이피항αvβ3、αvβ5적항체혹침묵β3화β5적소간우RNA(siRNA)조단.결론 세포점부분자αvβ3、αvβ5급기배체재종류세포-종류내피세포적점부과정중기중요개도작용.
Objective To investigate the role of adhesion molecules αvβ3 and αvβ5 and their ligands Del-1 and L1 in the tumor-endothelial cell adhesion in vitro.Methods The expression of αvβ3,αvβ5 and ICAM-1 in liver sinusoidal endothelial cells (LSEC) and liver cancer endothelial cells(T3A)cultured under normoxia or hypoxia were analyzed by RT-PCR and fluorescent activated cell sorter(FACS).The expression of Del-1 and L1 in six tumor cell lines under normoxia or hypoxia were analyzed by RT-PCR and Western blot,respectively.The adhesion of dye-labeled tumor cells and endothelial LSEC and T3A cells was measured by a fluorescence plate reader after their culture.Results The expression of αvβ3 and αvβ5 were higher in T3A cells than that in LSEC cells,and were upregulated under hypoxia,while the expression of ICAM-1 was lower in T3A cells than that in LSEC cells,and was upregulated under hypoxia only in LSEC.The expression of Del-1 and L1 molecules were obviously different in various tumor cell lines and were differentially regulated under hypoxia.The adhesion of tumor cells with Del-1 or L1 expression was higher in T3A cells than that in LSEC cells,and was significantly increased under hypoxia condition.Furthermore,the adhesion of tumor cells to T3A could be inhibited by antibodies against αvβ3 and αvβ5,or SiRNAs for β3 and β5.Conclusion αvβ3 and αvβ5 and their ligands Del-1 and L1 may play an important role in tumor cell migration.