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

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 and Hamet, P (2003) [3H]-thymidine labelling of DNA triggers apoptosis potentiated by E1A-adenoviral protein. Apoptosis, 8 (2). pp. 199-208. ISSN 1360-8185

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[(3)H]-thymidine is commonly used to analyze the accumulation of [(3)H]-labeled chromatin fragments in cells undergoing apoptosis. This study shows that [(3)H]-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 high-level [(3)H]-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 [(3)H]-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 [(3)H]-thymidine-treated VSMC-E1A were reduced by the pan-caspase 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 [(3)H]-thymidine-labeled DNA. They also demonstrate that [(3)H]-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.

Item Type: Article
Uncontrolled Keywords: adenovirus e1a proteins,amino acid chloromethyl ketones,animals,apoptosis,caspase 3,caspases,chromatin,culture media, serum-free,cyclic amp,dna,dogs,dose-response relationship, drug,microscopy, phase-contrast,rats,signal transduction,sodium-potassium-exchanging atpase,swine,thymidine
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Depositing User: Pure Connector
Date Deposited: 16 Dec 2014 10:06
Last Modified: 21 Oct 2022 00:23
URI: https://ueaeprints.uea.ac.uk/id/eprint/51552
DOI: 10.1023/A:1022931028235

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