Viral and cellular oncogene expression during progressive malignant transformation of SV40 transformed human fibroblasts

Charles Lewis Goolsby*, Marianne Steiner, Jeffrey Nemeth

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

In vitro investigation of the multistep neoplastic progression which occurs during transformation of human cells has been hindered by resistance of human cells to both immortalization and tumorigenicity (Mut. Res. 199; 273, 1988). Previously our laboratory established a cell line, HSF4‐T12, by transfection of normal human foreskin fibroblasts with the plasmid pSV3‐neo which contains the early genes of simian virus 40 (SV40). A multi‐step progression in karyotypic alterations and transformed phenotype occurred resulting in a neoplastic cell line that was immortal, transformed, and tumorigenic. We have examined changes in the SV40 proteins, large T (T‐antigen) and small t (t‐antigen) antigens, and in the cellular protein, p53, during progressive transformation of these cells. Total viral protein expression relative to total cellular protein increased following immortalization of HSF4‐T12 as did the ratio of T‐antigen to t‐antigen. Interestingly, no significant change in DNA content accompanied immortalization. However, during the progressive in vitro transformation of HSF4‐T12 which occurred primarily post‐immortalization, DNA index increased to 1.6 but only small additional increases in T‐antigen expression were seen. No consistent or critical role for t‐antigen in development of the tumorigenic phenotype was found in this system.

Original languageEnglish (US)
Pages (from-to)748-756
Number of pages9
JournalCytometry
Volume12
Issue number8
DOIs
StatePublished - Jan 1 1991

Keywords

  • Bivariate analysis
  • Flow cytometry
  • Human diploid fibroblast transformation
  • Immunofluorescence
  • In vitro transformation
  • Multistep neoplasia
  • Papovavirus transformation
  • Simian virus 40
  • Total cellular protein
  • Tumorigenicity

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

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