Dedifferentiation of human hepatocytes by extracellular matrix proteins in vitro: quantitative and qualitative investigation of cytokeratin 7, 8, 18, 19 and vimentin filaments

doi:10.1016/S0168-8278(98)80293-6

Journal of Hepatology

Volume 28, Issue 4, April 1998, Pages 677-690

https://doi.org/10.1016/S0168-8278(98)80293-6 Get rights and content

Abstract

Background/Aims: Liver cirrhosis and carcinogenesis are accompanied by an alteration in extracellular matrix material. Histological studies reveal upregulation of the intermediate filaments cytokeratins 8 and 18 and de novo synthesis of vimentin, and cytokeratin 7 or 19 in hepatocytes. The aim of this study was to investigate how these two processes are linked.

Methods: Human hepatocytes were seeded: (i) on the matrix components collagen I, IV, laminin, or fibronectin; (ii) on stoichiometrically different complete matrices, derived from human placenta (matrix I) or the Englebreth-Holm-Swarm tumor (matrix II), and (iii) inside a three-dimensional collagen I sandwich. Filament expression and assembly were measured by cytofluor analysis or confocal laserscan microscopy.

Results: The matrix components or complete matrices triggered enhancement of cytokeratins 8 and 18 and de novo synthesis of cytokeratins 7, 19 and vimentin in a characteristic way. Confocal images demonstrated a dense and uniform network of cytokeratin 18 in freshly isolated cells, which was “replaced” by a few, thick protein bundles within 20 days. Interestingly, newly synthesized cytokeratin 19 structurally resembled the cytokeratin 19 organization in biliary epithelial cells. Marked cytokeratin alterations could be partially prevented when hepatocytes were grown in a three-dimensional collagen sandwich.

Conclusions: Pathological alterations to the chemical composition, molecular structure, or spatial arrangement of the liver matrix lead to specific changes in the intermediate filament pattern in human hepatocytes. We assume that degradation of the matrix results in pathological alterations to the hepatocyte-receptor matrix-ligand ratio, followed by a switch from physiological to pathological cell-activation.

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Dedifferentiation of human hepatocytes by extracellular matrix proteins in vitro: quantitative and qualitative investigation of cytokeratin 7, 8, 18, 19 and vimentin filaments

Abstract

Cell

Exp Cell Res

Hum Pathol

Path Res Pract

Gastroenterology

Hepatology

Hum Pathol

Biochim Biophys Acta

Biochem Pharmacol

Biochem Pharmacol

Exp Cell Res

Hepatology

Blood Rev

Scand J Gastroenterol

Exp Cell Res

Exp Cell Res

Expression of cytokeratins in normal and diseased livers and in primary liver carcinomas

Arch Pathol Lab Med

Expression of intermediate filament proteins in fetal and adult human kidney: modulations of intermediate filament patterns during development and in damaged tissue

Lab Invest

Tumor-associated trypsin participates in cancer cell-mediated degradation of extracellular matrix

Cancer Res

Biochemical interactions of tumor cells with the basement membrane

Ann Rev Biochem

Diazepam metabolism in cultured hepatocytes from rat, rabbit, dog, guinea pig, and man

Drug Metabol Dispos

Hepatocytes may produce laminin in fibrotic liver and in primary culture

Hepatology

Growth of normal human hepatocytes in primary culture: effect of hormones and growth factors on DNA synthesis

Hepatology

Regulation of glycogen phosphorylase activity in isolated human hepatocytes

Hepatology

Interspecies variability in mitoxantrone metabolism using primary cultures of hepatocytes isolated from rat, rabbit and humans

Biochem Pharmacol

Isolation and long-term culture of human hepatocytes

Surgery

Keratinocyte growth factor and fibroblast growth factor action on DNA synthesis in rat and human hepatocytes: modulation by heparin

Exp Cell Res

Isolation and separation of human adult hepatocytes: yield and purity by using different methods

Zentralbl Chir