Stepwise transformation of astrocytes by simian virus 40 large T antigen and epidermal growth factor receptor overexpression

Phyllis S. Frisa, Elizabeth I. Walter, Lei Ling, Hsing Jien Kung, James W. Jacobberger

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We have investigated the transformed phenotype of neonatal mouse cortical astrocytes immortalized by retrovirus-mediated transfer of the SV40 large T antigen gene. Expression of T antigen was driven by the Moloney murine leukemia virus long terminal repeat. Cell lines were selected based on coexpression of neomycin resistance, which provides a selection method believed to be unbiased for transformation state. Astrocyte cell lines derived in this manner express T antigen over a relatively narrow range (~4- fold), are contact inhibited, are able to enter a quiescent state in the presence of growth factors, and do not readily form colonies in soft agar. Compared to mortal astrocytes, the population growth rate is increased 3- fold, saturation densities are 4-fold higher, and the genome is relatively unstable as measured by the presence of DNA-aneuploid stem lines and by changes in DNA ploidy over time. However, changes in transformation phenotype occur at a low rate, making the cell lines amenable to experimentation. Most often, the growth phenotype remained unchanged during months of culture. Transfection of an epidermal growth factor receptor (EGFR) gene was used to generate a subline that was conditionally transformed (colony formation in soft agar was dependent on transforming growth factor α). v-raf transfection was used to generate constitutive transformation. Thus, these cell lines appear to be excellent experimental models for progressive transformation. With them, untested hypotheses of brain tumor progression derived from human genetic studies may be tested experimentally.

Original languageEnglish
Pages (from-to)223-233
Number of pages11
JournalCell Growth and Differentiation
Volume7
Issue number2
Publication statusPublished - Feb 1 1996
Externally publishedYes

Fingerprint

Simian virus 40
Viral Tumor Antigens
Epidermal Growth Factor Receptor
Astrocytes
Cell Line
Phenotype
Agar
Transfection
Moloney murine leukemia virus
erbB-1 Genes
Polyomavirus Transforming Antigens
Neomycin
Terminal Repeat Sequences
Ploidies
Population Growth
DNA
Medical Genetics
Transforming Growth Factors
Aneuploidy
Retroviridae

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Stepwise transformation of astrocytes by simian virus 40 large T antigen and epidermal growth factor receptor overexpression. / Frisa, Phyllis S.; Walter, Elizabeth I.; Ling, Lei; Kung, Hsing Jien; Jacobberger, James W.

In: Cell Growth and Differentiation, Vol. 7, No. 2, 01.02.1996, p. 223-233.

Research output: Contribution to journalArticle

Frisa, Phyllis S. ; Walter, Elizabeth I. ; Ling, Lei ; Kung, Hsing Jien ; Jacobberger, James W. / Stepwise transformation of astrocytes by simian virus 40 large T antigen and epidermal growth factor receptor overexpression. In: Cell Growth and Differentiation. 1996 ; Vol. 7, No. 2. pp. 223-233.
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