Regulation of cell morphology and cytochrome P450 expression in human hepatocytes by extracellular matrix and cell-cell interactions
Regulation of cell morphology and cytochrome P450 expression in human hepatocytes by extracellular matrix and cell-cell interactions
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DOI：
10.1007/s004410100429
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发表时间：
2001-10
影响因子：
3.6
通讯作者：
G. Hamilton;Summer L Jolley;D. Gilbert;James D. Coon;Scott A. Barros;E. LeCluyse
中科院分区：
文献类型：
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作者：
G. Hamilton;Summer L Jolley;D. Gilbert;James D. Coon;Scott A. Barros;E. LeCluyse
在人肝细胞原代培养物中，研究了细胞外基质条件和接种密度对细胞结构和细胞色素P450 3A 4（CYP 3A 4）组成性和化学诱导表达的影响。组成型和药物诱导的微粒体CYP 3A 4表达在维持在复杂或简单基质（基质胶vs胶原蛋白，I型）或三明治构型（即，在两层细胞外基质之间），尽管每种条件表现出明显不同的形态学特性。然而，随着铺板密度从100%降低至25%，在维持在简单胶原基质上的肝细胞中观察到CYP 3A 4的组成型和诱导水平均呈密度依赖性降低。细胞形状和细胞结构的显着变化，同时注意到细胞间隙连接和E-钙粘蛋白介导的细胞粘附的表达和定位的减少。此外，随着细胞密度的降低，细胞内微管和微丝的分布也发生了显著变化，免疫反应性肌动蛋白和β-微管蛋白的表达也随之增加。这些影响在一定程度上逆转与细胞外基质覆盖单层或与另一种细胞类型的共培养。在低细胞密度下，使用Matrigel基质维持肝细胞中正常细胞形状和细胞骨架分布的努力未能恢复CYP 3A 4表达或对利福平（RIF）的反应性的正常基础水平。同样，E-钙粘蛋白和Cx-32表达再次减少，即使细胞骨架元素的分布和表达恢复到正常水平。这些结果表明，细胞-细胞接触，而不是细胞外基质的配置或组成，在确定正常的反应，在人类肝细胞的化学调节剂中发挥了关键作用。
The influence of extracellular matrix conditions and plating density on cell cytoarchitecture and the constitutive and chemically induced expression of cytochrome P450 3A4 (CYP3A4) was examined in primary cultures of human hepatocytes. Constitutive and drug-induced microsomal CYP3A4 expression occurred equally well in human hepatocyte cultures maintained on either a complex or simple substratum (Matrigel vs collagen, type I), or in a sandwich configuration (i.e., between two layers of extracellular matrix), despite the markedly different morphological properties exhibited by each condition. However, a density-dependent decrease in both the constitutive and induced levels of CYP3A4 was observed in hepatocytes maintained on a simple collagen substratum as plating density was reduced from 100% to 25%. Marked alterations in cell shape and cytoarchitecture were noted concomitant with decreases in the expression and localization of intercellular gap junctions and E-cadherin-mediated cell adhesions. In addition, the intracellular distribution of microtubules and microfilaments was altered substantially and the expression of immunoreactive actin and β-tubulin increased as cell density was decreased. These effects were reversed to some extent by overlaying monolayers with extracellular matrix or by co-culturing with another cell type. Efforts to maintain normal cell shape and cytoskeletal distribution in hepatocytes at low cell density with a Matrigel substratum failed to restore normal basal levels of CYP3A4 expression or responsiveness to rifampicin (RIF). Likewise, E-cadherin and Cx-32 expression was again reduced, even though the distribution and expression of cytoskeletal elements returned to normal levels. These results suggest that cell-cell contacts, but not the extracellular matrix configuration or composition, play a critical role in determining normal responsiveness to chemical modulators in human hepatocytes.