Expression of CCAAT/Enhancer Binding Protein Family Genes in Monolayer and Sandwich Culture of Hepatocytes: Induction of Stress-Inducible GADD153

Wilkinson, Roseanne C.; Dickson, Alan J.

doi:10.1006/bbrc.2001.6090

Cited by 5 publications

(5 citation statements)

References 40 publications

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“…Levels of caspase‐12 or the Bcl‐2 family (Bid, Bad, Bax, and Bak) were not significantly different in wild type and IRF‐1‐deficient hepatocytes. Recently, Wilkinson and Dickson reported that only stimulation by monolayer cultures induces CHOP/GADD153 expression in rat hepatocytes [Wilkinson and Dickson, 2001]. In fact, IFN‐γ enhanced expression at the mRNA level (Fig.…”

Section: Discussionmentioning

confidence: 96%

“…6A) and induced the protein in the nuclear fraction. There may be a variety of factors, such as molecular distribution, species differences, and medium and culture conditions, that regulate the expression of this molecule [Wilkinson and Dickson, 2001]. Furthermore, to establish whether CHOP/GADD153 is essential for IFN‐γ‐induced apoptosis in hepatocytes requires further experiments using CHOP/GADD153‐deficient hepatocytes.…”

Section: Discussionmentioning

confidence: 99%

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Interferon‐γ induces reactive oxygen species and endoplasmic reticulum stress at the hepatic apoptosis

Watanabe

Сузукі

Haruyama

et al. 2003

J of Cellular Biochemistry

103

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Interferon-gamma (IFN-gamma) induces cell-cycle arrest and p53-independent apoptosis in primary cultured hepatocytes. However, the detailed mechanism, including regulating molecules, is still unclear. In this study, we found that IFN-gamma induced generation of reactive oxygen species (ROS) in primary hepatocytes and that pyrrolidinedithiocarbamate (PDTC), an anti-oxidant reagent, completely suppressed IFN-gamma-induced hepatic apoptosis. PDTC blocked apoptosis downstream from IRF-1 and upstream from caspase activation, suggesting that the generation of ROS occurred between these stages. However, IFN-gamma also induced the generation of ROS in IRF-1-deficient hepatocytes, cells insensitive to IFN-gamma-induced apoptosis. Moreover, a general cyclooxygenase (COX) inhibitor, indomethacin (but not the cyclooxygenase 2-specific inhibitor, NS-398) also inhibited the apoptosis without blocking the generation of ROS. Both PDTC and indomethacin also blocked IFN-gamma-induced release of cytochrome c from mitochondria. These results suggest that ROS are not the only or sufficient mediators of IFN-gamma-induced hepatic apoptosis. In contrast, we also found that IFN-gamma induced endoplasmic reticulum (ER) stress proteins, CHOP/GADD153 and caspase 12, in wild-type primary hepatocytes, but induced only caspase 12 and not CHOP/GADD153 protein in IRF-1-deficient hepatocytes. These results suggest that IFN-gamma induces ER stress in primary hepatocytes. Both the ROS and ER stress induced by IFN-gamma may be complementary mediators that induce apoptosis in primary hepatocytes.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Interferon‐γ induces reactive oxygen species and endoplasmic reticulum stress at the hepatic apoptosis

Watanabe

Сузукі

Haruyama

et al. 2003

J of Cellular Biochemistry

103

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“…A low rate of liver -specific gene transcription within the first 24-48h of culture, affects liverspecific mRNA levels after a few days, and has been described to underly the gradual loss of liver-specific functions [11,424]. Restoration of cell polarity by overlaying collagen gels on hepatocyte monolayer cultures, and/or by promoting homophilic and heterophilic intercellular contacts and signaling, upregulates liver-specific functioning and gap junctional intercellular communcation, and stimulates the formation of structures resembling in vivo bile canaliculi for weeks and months [10,11,226,418,425]. Meanwhile, the mRNAs of ubiquitously-expressed genes such as GAPDH are destabilized [419].…”

Section: Hepatocyte Spreading and Liver-specific Functioningmentioning

confidence: 99%

Molecular Mechanisms Underlying the Dedifferentiation Process of Isolated Hepatocytes and Their Cultures

Elaut

Henkens²,

Papeleu³

et al. 2006

CDM

260

213

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Primary hepatocytes and their cultures are a simple but versatile, well-controlled, and relatively easy to handle in vitro system that is well-accepted for investigating xenobiotic biotransformation, enzyme induction and inhibition, and (biotransformation-mediated) hepatotoxicity. In addition, hepatocyte cultures have proven to be valuable tools in the study of liver physiology, viral hepatitis, and liver regeneration and are proposed as an alternative to orthotopic liver transplantation. It has been observed, however, that a number of liver-specific functions are progressively lost with time when hepatocytes are isolated and cultivated. These phenotypic changes are primarily the result of fundamental changes in gene expression concomitant with a diminished transcription of the relevant liver-specific genes, and can be interpreted as a 'dedifferentiation' of the isolated hepatocytes. Ischemia-reperfusion stress induced during the isolation process, disruption of the normal tissue architecture, as well as an adaptation to the in vitro environment are underlying factors and will be extensively discussed. A detailed description of the regulation of the hepatocyte phenotype in vivo in the first section of this review will help to understand the effect of these factors on hepatocyte gene expression. Although different approaches, mainly mimicking the in vivo hepatocyte environment, have been succesfully used to prevent or slow down the dedifferentiation of primary hepatocytes in monolayer culture, the ideal hepatocyte-based culture model, characterized by a long-term expression of hepatocyte-specific functions comparable to the in vivo level, does not exist at the moment. Consequently, alternative strategies should focus on the isolation procedure, during which dedifferentiation is already initiated. In addition, identification of the conditions needed for the full in vitro maturation of hepatic progenitor cells to quiescent, functional hepatocyte-like cells opens promising perspectives.

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“…In vitro attempts to mimic these interactions consisted of collagen coating of culture dishes or cultivating the cells in a sandwich configuration. Although hepatocyte attachment, viability [20,27,40] and functionality [41,42] were improved in this configuration, induction of the stress-responsive C/EBP homologue, GADD153 (growth arrest and DNA damage gene 153), could not be prevented [42]. In general, a complex ECM, rather than individual components, seems to be required for retaining the differentiated phenotype.…”

Section: Cell-extracellular Matrix Interactionsmentioning

confidence: 93%

“…However, in vitro, hepatocyte proliferation and differentiation exclude each other. A decrease in mRNA expression of the major hepatocyte-specific transcription factors, especially C/EBPα, has been observed already during the initial stages of culture [42,69]. In addition, the appearance of c-jun has been shown to precede the loss of total cytochrome P450 content [92].…”

Section: Isolation Of Hepatocytes: a Major Trigger For Dedifferentiationmentioning

confidence: 99%

Histone Deacetylase Inhibition: A Differentiation Therapy for Cultured Primary Hepatocytes?

Papeleu

Vanhaecke

Rogiers

2006

CEI

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Histone deacetylase (HDAC) inhibitors are now widely recognized as powerful anticancer agents. Unlike conventional chemotherapeutics, they not only inhibit tumor growth and survival but also reinitiate differentiation processes. Surprisingly, little attention has been paid so far to their capacity as differentiating agents for primary non-malignant cells. Dedifferentiation, however, is a common pernicious event occurring in almost all primary epithelial cell cultures and limits their use in fundamental and applied research. In this review, we elaborate on the innovative concept of HDAC inhibition as a key tool in the development of functional long-term cultures of primary hepatocytes. The need for long-term hepatocyte-based in vitro models is high and underscored by the fact that (sub)-chronic liver toxicity testing still consumes an important number of animals and by recent changes in EU legislation (e.g. chemicals policy REACH and cosmetics Directive 2003/15/EC). In addition, many cell-based therapies would benefit from the availability of highly differentiated primary hepatocytes as well. We believe that the HDAC inhibition approach could be applicable to create stable, differentiated culture systems of other primary cell types as well. In addition, HDAC inhibitors, in combination with tissue-specific growth factors, can be used to generate tissue-specific cell types from stem cells.

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Expression of CCAAT/Enhancer Binding Protein Family Genes in Monolayer and Sandwich Culture of Hepatocytes: Induction of Stress-Inducible GADD153

Cited by 5 publications

References 40 publications

Interferon‐γ induces reactive oxygen species and endoplasmic reticulum stress at the hepatic apoptosis

Interferon‐γ induces reactive oxygen species and endoplasmic reticulum stress at the hepatic apoptosis

Molecular Mechanisms Underlying the Dedifferentiation Process of Isolated Hepatocytes and Their Cultures

Histone Deacetylase Inhibition: A Differentiation Therapy for Cultured Primary Hepatocytes?

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