人海绵状血管瘤内皮细胞特性的体外研究

Characteristics of endothefial cells derived from human hepatic cavernous hemangioma in vitro

目的研究人海绵状血管瘤内皮细胞在形态、表型和功能方面的改变。方法从人海绵状血管瘤组织分离、纯化血管内皮细胞,并应用透射电子显微镜、流式细胞术、逆转录聚合酶链反应(RT-PCR)、明胶酶谱法和激光共聚焦显微镜等技术与人正常人肝窦内皮细胞进行比较分析。结果与正常人肝窦内皮细胞相比,人海绵状血管瘤内皮细胞的超微结构上,具有异常膨大的内质网和空泡样结构;表型上,高表达整合素αvβ3;功能上,体外形成血管样网络结构异常,并可能与其高表达的内源性血管内皮细胞生长因子、血管生成素1以及基质金属蛋白酶2有关。结论人海绵状血管瘤内皮细胞在形态、表型和功能特性上都不同于正常肝窦内皮细胞。

Objective To investigate the characteristics of endothelial cells derived from human cavernous hemangioma in morphology, phenotypes and functions, Methods Endothelial cells were isolated from human hepatic cavernous hemangioma. The morphological, and phenotypical and functional features of these cells were analyzed by transmission electron microscopy, fluorescence-activated cell sorter, RT-PCR, zymography,and confocal microscopy, Human liver sinusoidal endothelial cells （LSEC） were used as control, Results As compared with the LSEC, abnormally expanded endoplasmic reticulums and similarly arranged cytoplasmic vacuoles were found in the endothelial cells derived from hepatic cavernous hemangioma （HCHEC） by transmission electron microscopy, Flow cytometry showed that expression of αvβ3 was significantly increased in the HCHEC, The mRNA of vascular endothelial cell growth factor and angiopoietin 1 were more abundant in HCHEC than that in LSEC. Functional analysis indicated that the HCHEC exhibited strong activated angiogenesis capacity and formed abnormal capillary-like structures. HCHEC produced more pro-matrix metalloproteinase 2 （MMP-2） and the activated MMP-2 form as compared with the LSEC. Confocal microscopy revealed that MMP-2 was concentrated in those cytoplasmic granules of the HCHEC and was consistent with the distribution of the expanded endoplasmic reticulums, Conclusion The endothelial cells derived from human cavernous hemangioma differ from the normal endothelial cells in morphology, phenotypes and functions.