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

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5 引文 斯高帕斯(Scopus)


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.
頁(從 - 到)223-233
期刊Cell Growth and Differentiation
出版狀態已發佈 - 2月 1 1996

ASJC Scopus subject areas

  • 分子生物學
  • 細胞生物學


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