title: Extracellular matrix regulates the hepatocellular heat shock response.
abstract: BACKGROUND
  Cirrhosis is characterized by the accumulation of collagen within the extracellular matrix (ECM) of the liver and progressive hepatocellular dysfunction. Since recent studies have shown that the ECM can modulate cellular function, we examined whether the ECM could contribute to hepatocellular dysfunction. To address this question we examined hepatocyte behavior in two different ECM environments.
  
  
  METHODS
  Primary rat hepatocytes were cultured as a monolayer on collagen or as multicellular aggregates (spheroids) within a laminin-rich ECM. Hepatocytes were then compared for viability, response to proinflammatory cytokines, and their capacity to activate a heat shock response and adopt a thermotolerant phenotype. In addition, we compared the ability of prior heat shock exposure to protect hepatocytes from tumor necrosis factor (TNF) alpha/actinomycin-D-induced apoptosis in the two different ECM environments.
  
  
  RESULTS
  Hepatocytes cultured as a monolayer on collagen exhibited decreased viability, underwent spontaneous apoptosis, and displayed an attenuated cytokine-stimulated nitric oxide production compared to hepatocytes cultured as spheroids. In response to heat, hepatocytes in both ECM environments expressed inducible heat shock protein 70 (hsp72). But, only the hepatocyte spheroids exhibited thermotolerance in response to a subsequent thermal challenge. In contrast to previous reports, induction of the heat shock response failed to protect hepatocytes against TNFalpha-induced apoptosis.
  
  
  CONCLUSIONS
  These data demonstrate that the ECM can play an integral role in specific hepatocellular behaviors. Furthermore, the progressive deposition of collagen within the ECM, which is characteristic of fibrotic liver diseases, may directly contribute to the progressive hepatocellular dysfunction observed in cirrhosis. Hepatocellular viability, response to proinflammatory cytokines, heat shock response, and thermotolerance were all altered depending on the composition of the ECM. In contrast, TNFalpha-induced apoptosis was independent of the composition of the ECM.
authors.affiliation: UCSF Surgical Research Laboratory at San Francisco General Hospital, San Francisco, California 94110, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Gosnell
authors.forename: C B
authors.initials: CB
authors.lastname: Wong
authors.forename: Z L
authors.initials: ZL
authors.lastname: Kumwenda
authors.forename: W J
authors.initials: WJ
authors.lastname: Welch
authors.forename: H W
authors.initials: HW
authors.lastname: Harris
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D053938:DNA Fragmentation; D003609:Dactinomycin / Q000494:pharmacology; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D005355:Fibrosis / Q000378:metabolism / Q000473:pathology; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000378:metabolism; D018869:Heat-Shock Response / Q000502:physiology*; D007797:Laminin / Q000378:metabolism; D008099:Liver / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D007797:Laminin; D011500:Protein Synthesis Inhibitors; D014409:Tumor Necrosis Factor-alpha; D003609:Dactinomycin; D009569:Nitric Oxide; D003094:Collagen