[3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein

Serge N. Orlov, D. V. Pchejetski, S. D. Sarkissian, V. Adarichev, S. Taurin, A. V. Pshezhetsky, J. Tremblay, G. V. Maximov, D. DeBlois, M. R. Bennett, P. Hamet

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7 Citations (Scopus)

Abstract

[3H]-thymidine is commonly used to analyze the accumulation of [3H]-labeled chromatin fragments in cells undergoing apoptosis. This study shows that [3H]-thymidine incorporation within DNA is sufficient per se to inhibit growth and to induce apoptosis in canine kidney epithelial cells and porcine aorta endothelial cells. Despite highlevel [3H]-thymidine-DNA labeling, rat vascular smooth muscle cells (VSMC) showed only modest inhibition of growth and induction of apoptosis compared to other cell types. Similarly to serum deprivation, apoptosis triggered by [3H]-thymidine labeling was sharply potentiated by VSMC transfection with a functional analogue of c-myc, E1A-adenoviral protein (VSMC-E1A), and was suppressed by stimulation of cAMP signaling with forskolin as well as by and Na/K pump inhibition with ouabain. Both apoptosis induction and growth suppression seen in [3H]-thymidine-treated VSMC-E1A were reduced by the pancaspase inhibitor z-VAD.fmk. Thus, our results show that the differential efficiency of the apoptotic machinery determines cell type-specific attenuation of growth in cells with [3H]-thymidine-labeled DNA. They also demonstrate that [3H]-thymidine-treated and serum-deprived VSMC employ common intermediates of the apoptotic machinery, including steps that are potentiated by E1A-adenoviral protein and inhibited by activation of cAMP signaling as well as by inversion of the intracellular [Na+]i/[K+]i ratio.

Original languageEnglish
Pages (from-to)199-208
Number of pages10
JournalApoptosis
Volume8
Issue number2
DOIs
Publication statusPublished - Mar 1 2003

Keywords

  • Apoptosis
  • Cell growth
  • E1A adenoviral protein
  • [H]-thymidine

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science
  • Clinical Biochemistry
  • Cell Biology
  • Biochemistry, medical
  • Cancer Research

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  • Cite this

    Orlov, S. N., Pchejetski, D. V., Sarkissian, S. D., Adarichev, V., Taurin, S., Pshezhetsky, A. V., Tremblay, J., Maximov, G. V., DeBlois, D., Bennett, M. R., & Hamet, P. (2003). [3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein. Apoptosis, 8(2), 199-208. https://doi.org/10.1023/A:1022931028235