Transformation of differentiated neonatal rat hepatocytes in primary culture by polyoma virus early region sequences.
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Transformation of differentiated neonatal rat hepatocytes in primary culture by polyoma virus early region sequences. / Höhne, M; Piasecki, Angelika; Ummelmann, E; Paul, D.
in: ONCOGENE, Jahrgang 1, Nr. 4, 4, 1987, S. 337-345.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Transformation of differentiated neonatal rat hepatocytes in primary culture by polyoma virus early region sequences.
AU - Höhne, M
AU - Piasecki, Angelika
AU - Ummelmann, E
AU - Paul, D
PY - 1987
Y1 - 1987
N2 - Neonatal rat hepatocytes (NRH) in primary cultures were transformed upon transfection with plasmid pSV 5-neo containing polyoma virus (Py) early region sequences. Clones of cells (Py-NRH) resistant to the antibiotic G418 were obtained after selection in arginine-deficient medium with serum, epidermal growth factor (EGF), transferrin and insulin. They did not undergo senescence during serial subcultivation. One clone (Py-NRH Cl A) harbored a single integrated copy of Py early region sequences and expressed transforming Py genes, hepatocyte-specific transcripts, including albumin, alpha-fetoprotein (AFP) and tyrosine aminotransferase (TAT) mRNAs. Subclones isolated after about 45 cell doublings still contained albumin and AFP, but no TAT mRNAs, indicating that long-term stabilization of liver functions is not necessarily permanent unless selected for (e.g. arginine synthesis). Cells grew unrestricted in medium containing insulin and no longer required EGF. Cells grew in agar, secreted a beta-transforming growth factor-like activity into the medium and were tumorigenic in nude mice. Hybridization studies using v-erbB DNA as a probe showed that Py-NRH, unlike neonatal hepatocytes in primary culture, express the EGF receptor gene at low or undetectable levels. Py-NRH Cl A and a subclone (5A) derived from it, however, contained elevated levels of rat c-neu oncogene-related RNA, whereas levels in another subclone (3A) were low or undetectable. These findings demonstrate that a proto-oncogene was activated after transfection of hepatocytes with DNA tumor virus transforming genes. However, the expression of c-neu oncogene is not related to the maintenance of the transformed state.
AB - Neonatal rat hepatocytes (NRH) in primary cultures were transformed upon transfection with plasmid pSV 5-neo containing polyoma virus (Py) early region sequences. Clones of cells (Py-NRH) resistant to the antibiotic G418 were obtained after selection in arginine-deficient medium with serum, epidermal growth factor (EGF), transferrin and insulin. They did not undergo senescence during serial subcultivation. One clone (Py-NRH Cl A) harbored a single integrated copy of Py early region sequences and expressed transforming Py genes, hepatocyte-specific transcripts, including albumin, alpha-fetoprotein (AFP) and tyrosine aminotransferase (TAT) mRNAs. Subclones isolated after about 45 cell doublings still contained albumin and AFP, but no TAT mRNAs, indicating that long-term stabilization of liver functions is not necessarily permanent unless selected for (e.g. arginine synthesis). Cells grew unrestricted in medium containing insulin and no longer required EGF. Cells grew in agar, secreted a beta-transforming growth factor-like activity into the medium and were tumorigenic in nude mice. Hybridization studies using v-erbB DNA as a probe showed that Py-NRH, unlike neonatal hepatocytes in primary culture, express the EGF receptor gene at low or undetectable levels. Py-NRH Cl A and a subclone (5A) derived from it, however, contained elevated levels of rat c-neu oncogene-related RNA, whereas levels in another subclone (3A) were low or undetectable. These findings demonstrate that a proto-oncogene was activated after transfection of hepatocytes with DNA tumor virus transforming genes. However, the expression of c-neu oncogene is not related to the maintenance of the transformed state.
KW - Animals
KW - Gene Expression Regulation
KW - Cells, Cultured
KW - Rats
KW - Receptor, Epidermal Growth Factor/genetics
KW - Genes, Viral
KW - RNA, Messenger/genetics
KW - DNA, Viral/genetics
KW - Cell Transformation, Viral
KW - Liver/cytology/microbiology/physiology
KW - Peptide Biosynthesis
KW - Polyomavirus
KW - Proto-Oncogene Proteins/genetics
KW - RNA, Viral/genetics
KW - Recombination, Genetic
KW - Transforming Growth Factors
KW - Animals
KW - Gene Expression Regulation
KW - Cells, Cultured
KW - Rats
KW - Receptor, Epidermal Growth Factor/genetics
KW - Genes, Viral
KW - RNA, Messenger/genetics
KW - DNA, Viral/genetics
KW - Cell Transformation, Viral
KW - Liver/cytology/microbiology/physiology
KW - Peptide Biosynthesis
KW - Polyomavirus
KW - Proto-Oncogene Proteins/genetics
KW - RNA, Viral/genetics
KW - Recombination, Genetic
KW - Transforming Growth Factors
M3 - SCORING: Journal article
VL - 1
SP - 337
EP - 345
JO - ONCOGENE
JF - ONCOGENE
SN - 0950-9232
IS - 4
M1 - 4
ER -
