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Gene Expression in Clonally Derived Cell Lines Produced by in Vitro Transformation of Rat Fetal Hepatocytes: Isolation of Cell Lines Which Retain Liver-specific Markers¹

Department of Physiology, University of Western Australia, Nedlands, Western Australia 6009

The pattern of gene expression in fetal hepatocytes transformed in culture with a hepatocarcinogen (FRL cells) is studied with respect to a range of markers which are either developmentally regulated and/or shown to be expressed at high levels in hepatoma cells. The relative abundance of the respective mRNAs is determined and immunocytochemistry is used to detect the respective proteins in cultured cells. When compared with its normal counterpart, FRL cells retain the expression of transferrin, ₁-acid glycoprotein, -glutamyltranspeptidase, and tyrosine aminotransferase at near normal levels, while expression of the liver-specific isoenzymes of pyruvate kinase (L form) and aldolase (B form) is reduced. The cell lines are different in that they fail to express albumin, -fetoprotein, thiostatin and ₂-macroglobulin, and they express high levels of M₂-pyruvate kinase and aldolase A, markers often found in abundance in hepatoma cells. Therefore transformation has resulted in different effects on different genes. Furthermore, it is of interest to find that the cells coexpress both forms of the pyruvate kinase isoenzymes which does not occur in the normal developing hepatocyte.

These results indicate that it is possible to use this model to study changes which accompany transformation of fetal rat hepatocytes. The resulting cell lines have a stable phenotype and retain the changes which result from transformation even after extended passaging. This facilitates comparisons between the precursor cell and the tumor cell, both of which can be maintained under controlled conditions which exist in culture.

¹ This research was supported by grants from the National Health & Medical Research Council of Australia and The Faculty of Medicine, University of Western Australia.

Received 3/26/90. Accepted 8/ 4/90.

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