Cytosine arabinoside induces programmed endothelial cell death through the caspase-3 pathway

Ida M Moore, Carrie J Merkle, Petra Miketova, Renee K. Salyer, Bonny J. Torres, Richard C. Schaeffer, David W Montgomery

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The anti-cancer effects of cytosine arabinoside (ARA-C) are well known. However, effects on nonmalignant cells have not been elucidated and may be important to understanding treatment-related toxicity. The purpose of this study was to examine the effect of ARA-C on nondividing vascular endothelial cells. The objectives were to determine the effects of ARA-C on cell viability and to ascertain whether ARA-C caused apoptosis in cultured vascular endothelial cells and hydrocortisone blunted caspase-3-induced apoptosis. Endothelial cells were cultured until confluent and mitotically quiescent then exposed to ARA-C (10 -7 to 10 -3 M) for 1 to 4 days. Some experiments involved cotreatment with hydrocortisone (10 -11,10 -10,10 -4, and 10 -3 M). Light microscopy and the colorimetric MTS assay were used to measure viability. Fluorescent annexin-V and DNA fragmentation assays were used to measure apoptosis, and a fluorescence-based enzymatic assay was used to measure caspase-3 activity, which is one pathway involved in the apoptosis cascade. Two-way ANOVA or the appropriate nonparametric test was used to determine statistical significance in studies of viability and apoptosis. Oneway ANOVA was used to determine statistical significance for caspase-3 activity. Viability was decreased with higher concentrations of ARA-C and increased days of treatment. The percentage of apoptotic cells increased with higher concentrations of ARA-C and increased days of treatment. ARA-C-treated samples showed DNA fragmentation, indicative of apoptosis. Caspase-3 activity increased after ARA-C addition; hydrocortisone blunted this increase. ARA-C caused apoptosis in nondividing endothelial cells in culture. Hydrocortisone may protect against ARA-C-induced apoptosis by reducing caspase-3 activity.

Original languageEnglish (US)
Pages (from-to)289-296
Number of pages8
JournalBiological Research for Nursing
Volume7
Issue number4
DOIs
StatePublished - Apr 2006

Fingerprint

Cytarabine
Caspase 3
Cell Death
Endothelial Cells
Apoptosis
Hydrocortisone
DNA Fragmentation
Analysis of Variance
Annexin A5
Enzyme Assays
Microscopy
Cell Survival
Cell Culture Techniques
Fluorescence
Light

Keywords

  • Apoptosis
  • Caspase-3
  • Cell culture
  • Chemotherapy
  • Cytosine arabinoside
  • Drug toxicity
  • Endothelial cells
  • Microscopy

ASJC Scopus subject areas

  • Research and Theory

Cite this

Cytosine arabinoside induces programmed endothelial cell death through the caspase-3 pathway. / Moore, Ida M; Merkle, Carrie J; Miketova, Petra; Salyer, Renee K.; Torres, Bonny J.; Schaeffer, Richard C.; Montgomery, David W.

In: Biological Research for Nursing, Vol. 7, No. 4, 04.2006, p. 289-296.

Research output: Contribution to journalArticle

Moore, Ida M ; Merkle, Carrie J ; Miketova, Petra ; Salyer, Renee K. ; Torres, Bonny J. ; Schaeffer, Richard C. ; Montgomery, David W. / Cytosine arabinoside induces programmed endothelial cell death through the caspase-3 pathway. In: Biological Research for Nursing. 2006 ; Vol. 7, No. 4. pp. 289-296.
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AU - Montgomery, David W

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