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  • Title: Homocysteine increases the relative number of apoptotic cells and reduces the relative number of apoptotic bodies in HL-60 cells treated with 3-deazaadenosine.
    Author: Endresen PC, Prytz PS, Lysne S, Aarbakke J.
    Journal: J Pharmacol Exp Ther; 1994 Jun; 269(3):1245-53. PubMed ID: 8014868.
    Abstract:
    The effect of exogenous homocysteine thiolactone (Hcy) on apoptosis initiated by 3-deazaadenosine (c3Ado) was studied in human leukemia HL-60 cells. Flow cytometric analysis allowed evaluation of the relative number of apoptotic cells (APC) to apoptotic bodies (APB) with a sub G0/G1 DNA content. Addition of 1 mM Hcy to HL-60 cells exposed to 5 to 100 microM c3Ado was followed by a large increase in the number of APC and a simultaneous decrease in the number of APB. The effects of Hcy on both the formation of APC and APB was dose-dependent; however, Hcy concentrations above 250 microM were required for inhibition of APB formation to take place. Fluorescence microscopic examination of unfixed cells stained with acridine orange demonstrated different morphology of APC between cultures treated with c3Ado or c3Ado plus Hcy. Whereas APC in cultures treated with 100 microM c3Ado displayed pronounced cytoplasmic membrane blebbing, only minor blebbing was displayed by APC in cultures treated with 25 microM c3Ado and 1 mM Hcy. Extensive nuclear fragmentation was observed in APC regardless of Hcy addition. By cell sorting we demonstrate the presence of APC with the same DNA content as viable G0/G1 and S-phase cells in cultures treated with 25 microM c3Ado and 1 mM Hcy, indicating that cells in all cell cycle phases undergo apoptosis in these cultures. Neither the formation of APC nor APB in apoptosis initiated by cycloheximide or dactinomycin were influenced by 1 mM Hcy. The Hcy effects on c3Ado apoptosis were abrogated in part by 3-deaza-(+/-)-aristeromycin, a more specific and potent inhibitor of S-adenosylhomocysteine hydrolase.(ABSTRACT TRUNCATED AT 250 WORDS)
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