目的：研究人肝癌微血管内皮细胞(human liver cancer microvascular endothelial cell，HLCMVEC)上黏附分子表达的特点及其对免疫细胞黏附和迁移的影响。 方法： 分离培养HLCMVEC，以人肝窦内皮细胞系(liver sinusoid endothelial cell，LSEC)作为正常对照。通过细胞ELISA方法比较多种黏附分子在两种内皮细胞上的表达。外周血单个核细胞（peripheral blood mononuclear cell, PBMC）用荧光染料BCECF标记，将其与HLCMVEC或LSEC共培养，随后采用倒置荧光显微镜和连续光谱荧光仪检测PBMC在两种内皮细胞上的黏附和跨内皮迁移能力；此外，共培养前分别预加各种黏附分子的功能抗体，然后分析各种黏附分子在PBMC与肿瘤微血管内皮细胞黏附和迁移中的作用。 结果： HLCMVEC表达CD31、CD34和细胞间黏附分子-3 （in-tercellular adhesion molecule-3，ICAM-3）的水平低于LSEC(P<0.05)，表达ICAM-1和血管细胞黏附分子-1（vascular cell adhesion molecule-1，VCAM-1）的水平明显低于正常LSEC（P<0.01），但表达整合素αvβ3和αvβ5明显高于LSEC（P<0.01）。PBMC在HLC-MVEC上的黏附和趋化剂诱导下的跨内皮迁移均显著低于LSEC[黏附：(205.5±46.0) vs（330.5±48.4）个，迁移： （49.0±10.6）vs（110.0±19.2）个，均P<0.01]；该黏附和迁移可被ICAM-1、ICAM-3、VCAM-1抗体明显阻断（P<0.01），抗CD31抗体对黏附阻断不明显但能阻断跨内皮迁移（P<0.05）。 结论： HLCMVEC特有的黏附分子表达特点抑制了PBMC的黏附和跨内皮迁移。

Objective: To investigate the expression characteristics of adhesion molecules on human liver cancer microvascular endothelial cells (HLCMVECs) and its influence on adhesion and trans-endothelial migration of im-mune cells. Methods: HLCMVECs were isolated and cultured, and human liver sinusoid endothelial cells (LSECs) were used as control. The expression of adhesion molecules was evaluated by cell-based ELISA. Peripheral blood mononuclear cells (PBMCs) were labeled with BCECF (a fluorescent dye) and then co-cultured with HLCMVEC or LSEC; the adhesion to endothelial cells and trans-endothelial migration of PBMCs were observed by inverted fluo-rescence microscope and continuous spectrum luminoscope. In addition, the functional antibody against each adhe-sion molecule was respectively added to endothelial cells before co-culture, which could determine the contribution of each adhesion molecule to the adhesion and trans-endothelial migration of PBMC. Results: Compared to LSECs,HLCMVECs expressed low level of CD31, CD34 and intercellular adhesion molecule-3 (ICAM-3) (P<0.05) and an even lower level of intercellular adhesion molecule-1 (ICAM-1) and Vascular cell adhesion molecule-1 (VCAM-1)(P<0.01), but expressed significantly higher level of αvβ3 and αvβ5 (P<0.01). Compared with LSECs, the adhesion of PBMCs to HLVMVECs were significantly decreased; and under the inducement of chemo-tactic agent, trans-en-dothelial migration of PBMCs through HLVMVECs were also significantly decreased (adhesion: [205.5±46.0] vs [330.5±48.4]; migration: [49.0±10.6] vs [110.0±19.2]; all P<0.01). However, the adhesion and migration could be obviously blocked by antibodies against ICAM-3 (P<0.05), ICAM-1 and VCAM-1(P<0.01); Antibody against CD31 did not influence PBMC adhesion greatly but significantly reduced trans-endothelial migration (P<0.05).Conclusion: The distinct expression of adhesion molecules on HLCMVECs reduced the adhesion and trans-endo-thelial migration of PBMCs.

国家自然科学基金资助项目（No. 81402451, No. 81370873），国家重点基础研究发展计划（973 计划）资助项目（No.2009CB521804）