A plasma membrane protein is involved in cell contact-mediated regulation of tissue-specific genes in adult hepatocytes.

Corlu, Anne; Kneip, Bernard; Lhadi, C; Leray, G; Glaise, Denise; Baffet, Georges; Bourel, Dominique; Guguen‐Guillouzo, Christiane

doi:10.1083/jcb.115.2.505

Cited by 91 publications

(54 citation statements)

References 56 publications

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“…The mechanisms by which non-parenchymal cells allow sustained expression of differentiated functions of hepatocytes in co-cultures are not fully understood, although a key plasma membrane protein involved in hepatocyte-rat-liver-epithelialcell recognition in co-cultures has been identified (Corlu et al, 1991). Nevertheless, this culture model remains one of the most powerful and useful systems to study the effects of drug exposure over several weeks.…”

Section: Discussionmentioning

confidence: 99%

Regulation of the Major Detoxication Functions by Phenobarbital and 3‐Methylcholanthrene in Co‐Cultures of Rat Hepatocytes and Liver Epithelial Cells

Lerche

Fautrel

Shaw³

et al. 1997

European Journal of Biochemistry

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In the present study, we analysed the expression of monooxygenase activities and mRNAs associated with cytochrome P-450 (CUP), including CYPlAU2, CYP2B1/2, CYP2C6, CYP2E1, CYP3A1/2, glutathione transferase a (GSTa), aldehyde dehydrogenase and epoxide hydrolase in co-cultures of primary rat hepatocytes and rat liver epithelial cells. We observed that pentoxyresorufin 0-deethylation activity was well maintained and ethoxyresorufin O-deethylation activity gradually decreased during co-culture time. In addition, we showed that phenobarbital and 3-methylcholanthrene treatments resulted in a significant increase of these activities.Two general patterns of accumulation of liver-specific mRNAs were observed. CYPl A1 /2, CYP2B1/2, CYP3All2, GSTa, aldehyde dehydrogenase and epoxide hydrolase mRNAs were maintained at a stable level, whereas CYP2C6 and CYP2E1 mRNAs showed a continuous decline. In addition, we observed a strong increase of CYPl A1/2 (13.6-fold) and GSTu (3.9-fold) mRNA expression in 3-methylcholanthrene-treated co-cultures and induction of CYP2B1/2 (19-fold), CYP2C6 (10-fold), CYP3A1/2 (1 1 .Zfold), GSTu (9-fold), aldehyde dehydrogenase (6-fold) and epoxide hydrolase (5-fold) mRNA expression in phenobarbital-treated co-cultures. Furthermore, we demonstrated that liver-specific gene expression was restricted to hepatocytes, with the notable exception of epoxide hydrolase and CYP2E1 which were expressed in both cell types during the co-culture, as shown by the selective recovery of both hepatocytes and rat liver epithelial cells.Finally, to investigate whether co-cultures could be used to study the molecular mechanisms regulating CYP transcription, we performed transfection of hepatocytes, before the establishment of the co-culture, with large CYP2B 1 (3.9 kb) or CYP2B2 (4.5 kb) promoter chloramphenicol acetyltransferase constructs or with a construct containing a 163-bp DNA sequence element reported to confer phenobarbital responsiveness. A 2-3-fold increase over the basal level of chloramphenicol acetyltransferase activity was observed in phenobarbital-treated co-cultures transfected with the phenobarbital-responsive element construct, although phenobarbital had no effect on large CYP2B1 or CYP2B2 promoter fragments.Our results demonstrate that the co-culture system provides a good tool for studying drug metabolism, and shows promise as a new tool for analysing transcriptional regulation under the influence of xenobiotics within primary hepatocytes.

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Section: Discussionmentioning

confidence: 99%

Regulation of the Major Detoxication Functions by Phenobarbital and 3‐Methylcholanthrene in Co‐Cultures of Rat Hepatocytes and Liver Epithelial Cells

Lerche

Fautrel

Shaw³

et al. 1997

European Journal of Biochemistry

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“…Several studies have implicated cell surface proteins in epithelial-nonparenchymal interactions in hepatocyte cocultures. 17,18 Our gene expression profiling yielded Delta-like homolog 1, whose expression profile correlated positively (i.e., high-medium-low) with the ability of fibroblasts to induce functions in hepatocytes. Delta-like homolog 1 belongs to the epidermal growth factor-like homeotic protein family that includes proteins such as the Notch receptor and its homologues (see Table 2).…”

Section: Gene Expression Profiling Of Fibroblastsmentioning

confidence: 99%

“…14 Despite significant investigation, a complete picture of the molecular mediators of epithelial-nonparenchymal interaction in hepatocyte cocultures is unavailable. To date, the data suggest that both matrix deposition 15,16 and direct cell-cell contact play a role 10,17,18 in the "coculture effect," whereas soluble factors have proven to be largely ineffective. 14,19,20 Two promising candidate cell surface proteins are epithelial cadherin (E-cadherin) 17 and liverregulating protein 9 ; however, NPCs lacking E-cadherin and liver-regulating protein retain the ability to support hepatic functions, suggesting that neither is the sole mediator of the "coculture effect."…”

mentioning

confidence: 99%

Exploring interactions between rat hepatocytes and nonparenchymal cells using gene expression profiling

et al. 2004

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Cocultivation of primary hepatocytes with a plethora of nonparenchymal cells (from within and outside the liver) has been shown to support hepatic functions in vitro. Despite significant investigation into this phenomenon, the molecular mechanism underlying epithelial-nonparenchymal interactions in hepatocyte cocultures remains poorly understood. In this study, we present a functional genomic approach utilizing gene expression profiling to isolate molecular mediators potentially involved in induction of liver-specific functions by nonparenchymal cells. Specifically, primary rat hepatocytes were cocultivated with closely related murine fibroblast cell types (3T3-J2, NIH-3T3, mouse embryonic fibroblasts) to allow their classification as "high," "medium," or "low" inducers of hepatic functions. These functional responses were correlated with fibroblast gene expression profiles obtained using Affymetrix GeneChips. Microarray data analysis provided us with 17 functionally characterized candidate genes in the cell communication category (cell surface, extracellular matrix, secreted factors) that may be involved in induction of hepatic functions. Further analysis using various databases (i.e., PubMed, GenBank) facilitated prioritization of the candidates for functional characterization. We experimentally validated the potential role of two candidates in our coculture model. The cell surface protein, neural cadherin (N-cadherin), was localized to hepatocyte-fibroblast junctions, while adsorbed decorin up-regulated hepatic functions in pure cultures as well as cocultures with low-inducing fibroblasts. In the future, identifying mediators of hepatocyte differentiation may have implications for both fundamental hepatology and cell-based therapies (e.g., bioartificial liver devices). In conclusion, the functional genomic approach presented in this study may be utilized to investigate mechanisms of cell-cell interaction in a variety of tissues and disease states.

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“…Moreover, other cells have been effective, including both liver endothelial cells and nonhepatic cells (145)(146)(147). Studies from our laboratory (148,149) have shown that the presence of an integral plasma membrane glycoprotein is a prerequisite to obtain longterm survival of differentiated hepatocytes in coculture. This glycoprotein is expressed by both hepatocytes and rat liver epithelial cells and is also found in some other tissues (148,149).…”

Section: However Survival and Metabolic Compe-mentioning

confidence: 99%

“…Studies from our laboratory (148,149) have shown that the presence of an integral plasma membrane glycoprotein is a prerequisite to obtain longterm survival of differentiated hepatocytes in coculture. This glycoprotein is expressed by both hepatocytes and rat liver epithelial cells and is also found in some other tissues (148,149). One could postulate that nonhepatic cells that express this glycoprotein are potentially capable of interacting with hepatocytes.…”

Section: However Survival and Metabolic Compe-mentioning

confidence: 99%

Liver cell models in in vitro toxicology.

Guillouzo

1998

Environ Health Perspect

300

125

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In vitro liver preparations are increasingly used for the study of hepatotoxicity of chemicals. In recent years their actual advantages and limitations have been better defined. The cell models, slices, and mainly primary hepatocyte cultures, appear to be the most powerful in vitro systems, as liver-specific functions and responsiveness to inducers are retained either for a few days or several weeks depending on culture conditions. Maintenance of phase and phase 11 xenobiotic metabolizing enzyme activities allows various chemical investigations to be performed, including determination of kinetic parameters, metabolic profile, interspecies comparison, inhibition and induction effects, and drug-drug interactions. In vitro liver cell models also have various applications in toxicology: screening of cytotoxic and genotoxic compounds, evaluation of chemoprotective agents, and determination of characteristic liver lesions and associated biochemical mechanisms induced by toxic compounds. Extrapolation of the results to the in vivo situation remains a matter of debate. Presently, the most convincing applications of liver cell models are the studies on different aspects of metabolism and mechanisms of toxicity. For the future, there is a need for better culture conditions and differentiated hepatocyte cell lines to overcome the limited availability of human liver tissues. In addition, strategies for in vitro analysis of potentially toxic chemicals must be better defined. Environ Health Perspect 106(Suppl 2):511-532 (1998). http.//ehpnetl.niehs.nih.gov/docs/1998/Suppl-2/51 1-532guillouzo/abstract.html

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A plasma membrane protein is involved in cell contact-mediated regulation of tissue-specific genes in adult hepatocytes.

Cited by 91 publications

References 56 publications

Regulation of the Major Detoxication Functions by Phenobarbital and 3‐Methylcholanthrene in Co‐Cultures of Rat Hepatocytes and Liver Epithelial Cells

Regulation of the Major Detoxication Functions by Phenobarbital and 3‐Methylcholanthrene in Co‐Cultures of Rat Hepatocytes and Liver Epithelial Cells

Exploring interactions between rat hepatocytes and nonparenchymal cells using gene expression profiling

Liver cell models in in vitro toxicology.

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