Extended life span of human endometrial stromal cells transfected with cloned origin-defective, temperature-sensitive simian virus 40.
Human endometrial stromal cells transfected with an origin-defective, temperature-sensitive simian virus 40 recombinant plasmid are dependent on T-antigen function for proliferation and at the permissive temperature have an extended life span in culture. Southern blot analysis indicates that the transfected gene is present in low copy number, possibly at a single integration site. Normal stromal cells are capable of 10 to 20 population doublings in culture. Transfected cultures have been carried at the permissive temperature to 80 population doublings before crisis. In the multistep model of malignant transformation of human cells, these cells represent one of the earliest stages: extended but finite life span. We have used these cells to investigate alterations in signal transduction that may be responsible for this early stage of transformation caused by the large T antigen. Temperature shift experiments indicate that the expression of ornithine decarboxylase (ODC) but not of c-fos is altered by the large T antigen. Induction of c-fos by serum or 12-O-tetradecanoylphorbol-13-acetate is independent of temperature. However, in the transfected cells, the induction of ODC by asparagine or serum is greatly enhanced at the permissive temperature. This result indicates that the large T antigen acts downstream of c-fos but upstream of ODC expression in the signal-transducing cascade.
Rinehart
C A
CA
Department of Pathology, University of North Carolina, Chapel Hill 27599-7525.
Haskill
J S
JS
Morris
J S
JS
Butler
T D
TD
Kaufman
D G
DG
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002470:Cell Survival; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004717:Endometrium / Q000166:cytology*; D005260:Female; D006801:Humans; D007700:Kinetics; D009955:Ornithine Decarboxylase / Q000096:biosynthesis / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016760:Proto-Oncogene Proteins c-fos; D011519:Proto-Oncogenes; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D015398:Signal Transduction; D013539:Simian virus 40 / Q000235:genetics*; D013696:Temperature; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D012367:RNA, Viral; D011505:Protein-Tyrosine Kinases; D009955:Ornithine Decarboxylase; D013755:Tetradecanoylphorbol Acetate