Cajero-Juárez Marcos, Avila Bibiana, Ochoa Alejandra, Garrido-Guerrero Efraín, Varela-Echavarría Alfredo, Martínez de la Escalera Gonzalo, Clapp Carmen
Neurobiology Center, National Autonomous University of México, Querétaro.
Eur J Cell Biol. 2002 Jan;81(1):1-8. doi: 10.1078/0171-9335-00213.
Endothelial cells perform a large array of physiological functions that are influenced by their cellular heterogeneity in the different vascular beds. Vein endothelial cells isolated from the umbilical cords are commonly used to study vascular endothelium. Primary cultures of these cells, however, have low proliferative capacity and a limited life span. We have immortalized bovine umbilical vein endothelial cells (BUVEC) by transfection with an expression vector containing the human papillomavirus type 16 E6E7 oncogenes. Expression of E6E7 extended the life span of BUVEC from 40 to more than 1-20 cell replication cycles with no signs of senescence. Four immortalized clones were isolated and found to maintain endothelial cell properties, such as the uptake of acetylated low density lipoprotein, the expression of the von Willebrand protein, the binding of endothelial cell-specific lectins and proliferative responses to the specific endothelial cell mitogen, vascular endothelial growth factor. Moreover, clone BVE-E6E7-1, like its wild-type counterparts, expressed prolactin mRNA and decreased its proliferation in response to the anti-angiogenic 16-kDa fragment of prolactin. This clone showed little signs of genetic instability as revealed by centrosome and chromosome number analysis. Thus, immortalized E6E7 BUVEC cell lines retain endothelial cell characteristics and could facilitate studies to investigate the action of regulatory factors of vascular endothelium. Moreover, being the first non-human umbilical vein endothelial cell lines, their use should provide insights into the mechanisms governing species-related heterogeneity of endothelial cells.
内皮细胞执行大量生理功能，这些功能受其在不同血管床中的细胞异质性影响。从脐带分离的静脉内皮细胞常用于研究血管内皮。然而，这些细胞的原代培养物增殖能力低且寿命有限。我们通过用含有16型人乳头瘤病毒E6E7癌基因的表达载体转染，使牛脐静脉内皮细胞（BUVEC）永生化。E6E7的表达将BUVEC的寿命从40个细胞复制周期延长至超过120个细胞复制周期，且无衰老迹象。分离出四个永生化克隆，发现它们保持内皮细胞特性，如摄取乙酰化低密度脂蛋白、表达血管性血友病因子、结合内皮细胞特异性凝集素以及对特异性内皮细胞促分裂原血管内皮生长因子的增殖反应。此外，克隆BVE-E6E7-1与其野生型对应物一样，表达催乳素mRNA，并在对催乳素的抗血管生成16 kDa片段的反应中降低其增殖。如通过中心体和染色体数目分析所揭示，该克隆几乎没有遗传不稳定的迹象。因此，永生化的E6E7 BUVEC细胞系保留内皮细胞特征，可促进研究血管内皮调节因子的作用。此外，作为首批非人类脐静脉内皮细胞系，它们的使用应为了解控制内皮细胞物种相关异质性的机制提供见解。
