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Basic Sciences

Differential Cytokeratin and α-Fetoprotein Expression in Morphologically Distinct Epithelial Cells Emerging at the Early Stage of Rat Hepatocarcinogenesis

Lucie Germain, René Goyette and Normand Marceau
Lucie Germain
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René Goyette
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Normand Marceau
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DOI:  Published February 1985
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Abstract

The various liver cell populations emerging during the transitory reappearance of α-fetoprotein (AFP) in serum of 3′-methyl-4-dimethylaminoazobenzene-ingesting rats were analyzed in situ and in vitro on isolated cell preparations in terms of their cytokeratin and AFP expression using single and double indirect immunofluorescence microscopy. A polyclonal guinea pig antibody raised against cow hoof prekeratin, which recognized a Mr 52,000 cytokeratin, was found to react with bile ductular epithelial cells and oval cells but not with hepatocytes. A monoclonal antibody against a Mr 55,000 cytokeratin reacted not only with bile ductular and oval cells but also with hepatocytes. In contrast, a polyclonal antibody against porcine eye lens vimentin reacted with sinusoidal cells and stroma cells. To assess further the heterogeneity of the emerging cell populations, liver cells were isolated after 4 weeks of treatment and fractionated according to cell size and ploidy level into 4 fractions (I to IV) by velocity sedimentation at 1 × g. A cell-type analysis using AFP and albumin as functional markers revealed the presence of AFP-producing cells in Fraction IV at a mean velocity equivalent to that of newborn diploid rat hepatocytes, wheras most of the albumin-producing cells were distributed in Fractions I to III at velocities similar to those of adult tetraploid rat hepatocytes. A similar analysis based on the differential expression of Mr 52,000 and Mr 55,000 cytokeratins and vimentin in bile ductular and other diploid epithelial cells, hepatocytes, and mesenchymal cells showed that large cells in Fractions I to III were tetraploid hepatocytes, whereas viable cells present in Fraction IV were diploid epithelial cells and mesenchymal cells in proportion of 62 and 38%, respectively. These cell populations could be resolved further by changing the sedimentation time. A subsequent examination of the Mr55,000 cytokeratin-containing diploid epithelial cells in Fraction IV using double immunofluorescence microscopy resolved three cell populations with respect to Mr 52,000 cytokeratin and AFP expression, namely, two cell populations expressing either protein marker and a third one containing both markers. These results suggest a ductular origin of oval cells and a possible relation to immature hepatocytes.

Footnotes

  • ↵1 This investigation was supported by the National Cancer Institute of Canada.

  • ↵2 Recipient of a studentship from the Medical Research Council of Canada.

  • ↵3 To whom requests for reprints should be addressed, at Cancer Research Center, l'Hôtel-Dieu de Québec Hospital, 11 Côte du Palais, Québec City, Québec, Canada, G1R 2J6.

  • Received December 27, 1983.
  • Accepted October 30, 1984.
  • ©1985 American Association for Cancer Research.
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February 1985
Volume 45, Issue 2
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Differential Cytokeratin and α-Fetoprotein Expression in Morphologically Distinct Epithelial Cells Emerging at the Early Stage of Rat Hepatocarcinogenesis
Lucie Germain, René Goyette and Normand Marceau
Cancer Res February 1 1985 (45) (2) 673-681;

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Differential Cytokeratin and α-Fetoprotein Expression in Morphologically Distinct Epithelial Cells Emerging at the Early Stage of Rat Hepatocarcinogenesis
Lucie Germain, René Goyette and Normand Marceau
Cancer Res February 1 1985 (45) (2) 673-681;
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