E-cadherin protects primary hepatocyte spheroids from cell death by a caspase-independent mechanism.
E-cadherin protects primary hepatocyte spheroids from cell death by a caspase-independent mechanism.
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DOI：
10.3727/096368909x474258
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发表时间：
2009
影响因子：
3.3
通讯作者：
Nyberg SL
中科院分区：
文献类型：
--
作者：
Luebke-Wheeler JL;Nedredal G;Yee L;Amiot BP;Nyberg SL
将原代肝细胞培养成球状体为体外肝脏研究建立了一个有效的三维模型系统，并可作为球状贮库型生物人工肝的细胞来源。球状体形成的机制知之甚少，这也解释了为什么正常的锚定依赖性肝细胞在置于悬浮球状体培养物中时仍能存活，并且不发生凋亡诱导的细胞凋亡（称为失巢凋亡）。本研究旨在探讨钙依赖性细胞粘附分子E-钙粘蛋白在肝细胞球体形成和维持中的作用。通过一种新的摇杆技术形成肝细胞球状体，并在悬浮液中培养长达24小时。使用E-钙粘蛋白阻断抗体和钙螯合剂建立了球体形成对E-钙粘蛋白和钙的依赖性。我们发现，抑制E-钙粘蛋白阻止细胞-细胞附着和球体形成，令人惊讶的是，E-钙粘蛋白抑制导致肝细胞死亡，通过半胱天冬酶独立的机制。总之，E-cadherin是肝细胞球体形成所必需的，并且可能负责保护肝细胞免于一种新形式的半胱天冬酶非依赖性细胞死亡。
Cultivation of primary hepatocytes as spheroids creates an efficient three-dimensional model system for hepatic studies in vitro and as a cell source for a spheroid reservoir bioartificial liver. The mechanism of spheroid formation is poorly understood, as is an explanation for why normal, anchorage-dependent hepatocytes remain viable and do not undergo detachment-induced apoptosis, known as anoikis, when placed in suspension spheroid culture. The purpose of this study was to investigate the role of E-cadherin, a calcium-dependent cell adhesion molecule, in the formation and maintenance of hepatocyte spheroids. Hepatocyte spheroids were formed by a novel rocker technique and cultured in suspension for up to 24 h. The dependence of spheroid formation on E-cadherin and calcium was established using an E-cadherin blocking antibody and a calcium chelator. We found that inhibiting E-cadherin prevented cell–cell attachment and spheroid formation, and, surprisingly, E-cadherin inhibition led to hepatocyte death through a caspase-independent mechanism. In conclusion, E-cadherin is required for hepatocyte spheroid formation and may be responsible for protecting hepatocytes from a novel form of caspase-independent cell death.