Differential expression and distribution of focal adhesion and cell adhesion molecules in rat hepatocyte differentiation
Differential expression and distribution of focal adhesion and cell adhesion molecules in rat hepatocyte differentiation
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DOI：
10.1006/excr.1998.4209
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发表时间：
1998-10-10
影响因子：
3.7
通讯作者：
Michalopoulos, GK
中科院分区：
文献类型：
--
作者：
Kim, TH;Bowen, WC;Michalopoulos, GK
原代培养的肝细胞在含有肝细胞生长因子和表皮生长因子的化学限定的肝细胞生长介质中进入克隆性增殖。肝细胞增殖与分化基因表达缺失有关。来自Engelbreth-Holm-Spot小鼠肉瘤(Matrigel)的基质覆盖在增殖中的肝细胞上，诱导肝脏分化标志物基因的重新表达。为了探讨基质在肝细胞分化过程中的作用，我们检测了增殖肝细胞和Matrigel处理的肝细胞中纤维连接蛋白、玻璃体连接蛋白和肌动蛋白的mRNAs。纤维连接蛋白在第2~10天在增殖期肝细胞中表达增加，然后下降，而在增殖期肝细胞中消失，在Matrigel处理的肝细胞中重新表达。我们还发现Matrigel处理的肝细胞中粘着斑激酶和Paxlin显著增加，而E-钙粘附素和β-连环蛋白略有增加，这表明细胞与细胞外基质和细胞与细胞之间的相互作用可能是肝细胞分化的重要组成部分。为了评估局部黏附相关分子和细胞间黏附分子的分布，在第8天、第9天、第10天和第11天检测了Triton X-100在增殖肝细胞和Matrigel处理细胞中的可溶性和不溶性部分。我们发现，在Matrigel处理的肝细胞中，Triton X-100不溶性部分中的E-钙粘附素显著减少；而在Matrigel处理的肝细胞中，Triton X-100可溶部分中的β-catenin显著增加。这些结果表明，在Matrigel的覆盖诱导分化过程中，局部黏附相关分子和细胞黏附分子的分布都发生了重组。(C)1998年学术出版社。
Hepatocytes in primary culture enter into clonal proliferation in the chemically defined hepatocyte growth medium in the presence of hepatocyte growth factor and epidermal growth factor. Hepatocyte proliferation is associated with loss of differentiated gene expression. Overlay of matrix derived from Engelbreth-Holm-Swarm mouse sarcoma (Matrigel) on proliferating hepatocytes Hepatocytes induces reexpression of the hepatic differentiation marker genes. To explore the role of matrix in the differentiation process of hepatocytes, we examined the mRNAs of fibronectin, vitronectin, and entactin in proliferating hepatocytes and Matrigel-treated hepatocytes. Fibronectin mRNA increased in proliferating hepatocytes at days 2-10 and then decreased; however, vitronectin mRNA disappeared in proliferating hepatocytes and was reexpressed in Matrigel-treated hepatocytes. We also found that focal adhesion kinase and paxillin were strongly increased in Matrigel-treated hepatocytes, and E-cadherin and beta-catenin slightly increased in Matrigel-treated hepatocytes, suggesting that both cell-to extracellular matrix and cell-to-cell interactions may be an essential part of hepatocyte differentiation. To evaluate the distribution of focal adhesion associated molecules and cell-to-cell adhesion molecules, Triton X-100 soluble and insoluble fractions were examined at days 8, 9, 10, and 11 in proliferating hepatocytes and Matrigel-treated cells. We found that E-cadherin in Triton X-100 insoluble fractions dramatically decreased in Matrigel-treated hepatocytes; however, beta-catenin strongly increased in Triton X-100 soluble fractions of Matrigel-treated hepatocytes. These results suggest that the distribution of both focal adhesion associated molecules and cell adhesion molecules are reorganized during the process of differentiation induced by overlay of Matrigel. (C) 1998 Academic Press.