Mature vascular endothelium can give rise to smooth muscle cells via endothelial-mesenchymal transdifferentiation - In vitro analysis
Mature vascular endothelium can give rise to smooth muscle cells via endothelial-mesenchymal transdifferentiation - In vitro analysis
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DOI：
10.1161/01.res.0000021432.70309.28
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发表时间：
2002-06-14
影响因子：
20.1
通讯作者：
Stenmark, KR
中科院分区：
文献类型：
--
作者：
Frid, MG;Kale, VA;Stenmark, KR
虽然在过去被认为是一种罕见的现象，内皮间充质转分化已被描述与近年来越来越频繁。它被认为在胚胎血管发育中很重要，但对其在成人血管系统中的作用知之甚少。使用流式细胞仪和免疫磁性（Dynabeads）纯化技术（基于DiI-乙酰化低密度脂蛋白的摄取和/或PECAM-1表达）和双标记间接免疫染色（对于内皮和平滑肌[SM]标记物），我们证明成熟的牛血管内皮含有内皮表型的细胞，（由VE-钙粘蛋白，血管性血友病因子，PECAM-1，乙酰化低密度脂蛋白的摄取增加），其可以经历内皮-间充质转分化并进一步分化为SM细胞（如通过α-SM-肌动蛋白、SM 22 α、钙调蛋白和SM-肌球蛋白的表达所定义）。“过渡”细胞，共表达内皮标志物和α-SM-肌动蛋白，一贯观察。在原代内皮培养物中能够内皮-间充质转分化的细胞的百分比估计为0.01%至0.03%。即使在没有增殖的情况下，在γ-照射（30戈伊）和/或丝裂霉素C处理的原代细胞培养物中也发生SM表型的获得。转分化的启动与细胞-细胞接触的破坏相关（以VE-钙粘蛋白表达的丧失为标志），以及转化生长因子-β（1）的作用，总之，我们的体外数据显示，成熟的牛体和肺内皮细胞含有能够通过转化生长因子β（1）-和细胞-细胞接触依赖性，但不依赖于增殖。
Though in the past believed to be a rare phenomenon, endothelial-mesenchymal transdifferentiation has been described with increasing frequency in recent years. It is believed to be important in embryonic vascular development, yet less is known regarding its role in the adult vasculature. Using FACS and immunomagnetic (Dynabeads) purification techniques (based on uptake of DiI-acetylated low-density lipoproteins and/or PECAM-1 expression) and double-label indirect immunostaining (for endothelial and smooth muscle [SM] markers), we demonstrate that mature bovine vascular endothelium contains cells of an endothelial phenotype (defined by VE-cadherin, von Willebrand factor, PECAM-1, and elevated uptake of acetylated low-density lipoproteins) that can undergo endothelial-mesenchymal transdifferentiation and further differentiate into SM cells (as defined by expression of alpha-SM-actin, SM22alpha, calponin, and SM-myosin). "Transitional" cells, coexpressing both endothelial markers and alpha-SM-actin, were consistently observed. The percentage of cells capable of endothelial-mesenchymal transdifferentiation within primary endothelial cultures was estimated as 0.01% to 0.03%. Acquisition of a SM phenotype occurred even in the absence of proliferation, in gamma-irradiated (30 Gy) and/or mitomycin C-treated primary cell cultures. Initiation of transdifferentiation correlated with disruption of cell-cell contacts (marked by loss of VE-cadherin expression) within endothelial monolayers, as well as with the action of transforming growth factor-beta(1), In conclusion, our in vitro data show that mature bovine systemic and pulmonary endothelium contains cells that can acquire a SM phenotype via a transdifferentiation process that is transforming growth factor-beta(1)- and cell-cell contact-dependent, but proliferation-independent.