[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

Research output: Contribution to journalArticle

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 2003
Externally publishedYes

Fingerprint

Thymidine
Labeling
Vascular Smooth Muscle
Apoptosis
Smooth Muscle Myocytes
Muscle
DNA
Proteins
Growth
Machinery
Cells
Endothelial cells
Colforsin
Ouabain
Serum
Chromatin
Transfection
Aorta
Canidae
Rats

Keywords

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

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Cell Biology

Cite this

Orlov, S. N., Pchejetski, D. V., Sarkissian, S. D., Adarichev, V., Taurin, S., Pshezhetsky, A. V., ... 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

[3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein. / Orlov, Serge 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.

In: Apoptosis, Vol. 8, No. 2, 03.2003, p. 199-208.

Research output: Contribution to journalArticle

Orlov, SN, Pchejetski, DV, Sarkissian, SD, Adarichev, V, Taurin, S, Pshezhetsky, AV, Tremblay, J, Maximov, GV, DeBlois, D, Bennett, MR & Hamet, P 2003, '[3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein', Apoptosis, vol. 8, no. 2, pp. 199-208. https://doi.org/10.1023/A:1022931028235
Orlov SN, Pchejetski DV, Sarkissian SD, Adarichev V, Taurin S, Pshezhetsky AV et al. [3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein. Apoptosis. 2003 Mar;8(2):199-208. https://doi.org/10.1023/A:1022931028235
Orlov, Serge 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. / [3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein. In: Apoptosis. 2003 ; Vol. 8, No. 2. pp. 199-208.
@article{f1e3ff4c7a1a4a0eae5068a875fa0273,
title = "[3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein",
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.",
keywords = "[H]-thymidine, Apoptosis, Cell growth, E1A adenoviral protein",
author = "Orlov, {Serge N.} and Pchejetski, {D. V.} and Sarkissian, {S. D.} and V. Adarichev and S. Taurin and Pshezhetsky, {A. V.} and J. Tremblay and Maximov, {G. V.} and D. DeBlois and Bennett, {M. R.} and P. Hamet",
year = "2003",
month = "3",
doi = "10.1023/A:1022931028235",
language = "English",
volume = "8",
pages = "199--208",
journal = "Apoptosis : an international journal on programmed cell death",
issn = "1360-8185",
publisher = "Springer Netherlands",
number = "2",

}

TY - JOUR

T1 - [3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein

AU - Orlov, Serge N.

AU - Pchejetski, D. V.

AU - Sarkissian, S. D.

AU - Adarichev, V.

AU - Taurin, S.

AU - Pshezhetsky, A. V.

AU - Tremblay, J.

AU - Maximov, G. V.

AU - DeBlois, D.

AU - Bennett, M. R.

AU - Hamet, P.

PY - 2003/3

Y1 - 2003/3

N2 - [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.

AB - [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.

KW - [H]-thymidine

KW - Apoptosis

KW - Cell growth

KW - E1A adenoviral protein

UR - http://www.scopus.com/inward/record.url?scp=0037697865&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037697865&partnerID=8YFLogxK

U2 - 10.1023/A:1022931028235

DO - 10.1023/A:1022931028235

M3 - Article

C2 - 12766480

AN - SCOPUS:0037697865

VL - 8

SP - 199

EP - 208

JO - Apoptosis : an international journal on programmed cell death

JF - Apoptosis : an international journal on programmed cell death

SN - 1360-8185

IS - 2

ER -