Rho kinase, myosin-II, and p42/44 MAPK control extracellular matrix-mediated apical bile canalicular lumen morphogenesis in HepG2 cells
Rho kinase, myosin-II, and p42/44 MAPK control extracellular matrix-mediated apical bile canalicular lumen morphogenesis in HepG2 cells
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DOI：
10.1091/mbc.e06-01-0067
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发表时间：
2006-07-01
影响因子：
3.3
通讯作者：
van Ijzendoorn, Sven C. D.
中科院分区：
文献类型：
--
作者：
Herrema, Hilde;Czajkowska, Dorninika;van Ijzendoorn, Sven C. D.
调节肝细胞多细胞结构和扩展的顶端胆小管腔的发展的分子机制知之甚少。在这里，我们表明，肝HepG 2细胞培养的玻璃盖片上首先发展细胞间的顶端管腔通常形成的一对细胞。延长细胞培养导致广泛的组织变化，包括细胞聚集，多层和顶腔形态发生。后者包括大腺泡结构的发育和随后跨越多个细胞的细长小管腔。这些形态学变化与肝脏发育、再生和肿瘤形成过程中的早期组织模式非常相似，并且当细胞在预沉积的细胞外基质（ECM）上培养时迅速诱导。在盖玻片上培养的细胞中抑制Rho激酶或其靶肌球蛋白-II ATP酶模拟对ECM的形态发生反应。一致的是，刺激Rho激酶和随后的肌球蛋白-II ATP酶活性的脂氧合酶控制的二十碳四烯酸代谢抑制ECM介导的细胞多层和顶腔形态发生，但不初始顶腔形成。此外，顶腔重塑而不是细胞多层化需要基础p42/44 MAPK活性。总之，这些数据表明肝细胞来源的ECM在肝细胞和顶腔形态发生的空间组织中的作用，并确定Rho激酶，肌球蛋白-II和MAPK在胆管腔形态发生的不同方面可能是重要的参与者。
The molecular mechanisms that regulate multicellular architecture and the development of extended apical bile canalicular lumens in hepatocytes are poorly understood. Here, we show that hepatic HepG2 cells cultured on glass coverslips first develop intercellular apical lumens typically formed by a pair of cells. Prolonged cell culture results in extensive organizational changes, including cell clustering, multilayering, and apical lumen morphogenesis. The latter includes the development of large acinar structures and subsequent elongated canalicular lumens that span multiple cells. These morphological changes closely resemble the early organizational pattern during development, regeneration, and neoplasia of the liver and are rapidly induced when cells are cultured on predeposited extracellular matrix (ECM). Inhibition of Rho kinase or its target myosin-II ATPase in cells cultured on glass coverslips mimics the morphogenic response to ECM. Consistently, stimulation of Rho kinase and subsequent myosin-II ATPase activity by lipoxygenase-controlled eicosatetranoic acid metabolism inhibits ECM-mediated cell multilayering and apical lumen morphogenesis but not initial apical lumen formation. Furthermore, apical lumen remodeling but not cell multilayering requires basal p42/44 MAPK activity. Together, the data suggest a role for hepatocyte-derived ECM in the spatial organization of hepatocytes and apical lumen morphogenesis and identify Rho kinase, myosin-II, and MAPK as potentially important players in different aspects of bile canalicular lumen morphogenesis.