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Biological Research For Nursing
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*CYTARABINE
*HYDROCORTISONE
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Cytosine Arabinoside Induces Programmed Endothelial Cell Death Through the Caspase-3 Pathway

Ida M. (Ki) Moore, DNS, RN, FAAN

College of Nursing at the University of Arizona, Tucson, Southern Arizona VA Health Care System, Tucson, kmoore{at}nursing.arizona.edu

Carrie J. Merkle, RN, PhD, FAAN

College of Nursing at the University of Arizona, Tucson, Southern Arizona VA Health Care System, Tucson

Petra Miketova, PhD

College of Nursing at the University of Arizona, Tucson

Renee K. Salyer

College of Nursing at the University of Arizona, Tucson

Bonny J. Torres, BSN, RN

College of Nursing at the University of Arizona, Tucson

Richard C. Schaeffer, Jr

Southern Arizona VA Health Care System, Tucson

David W. Montgomery, PhD

College of Medicine at the University of Arizona, Tucson, Southern Arizona VA Health Care System, Tucson

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.

Key Words: cytosine arabinoside • apoptosis • endothelial cells • cell culture • chemotherapy • microscopy • drug toxicity • caspase-3

Biological Research For Nursing, Vol. 7, No. 4, 289-296 (2006)
DOI: 10.1177/1099800405286138
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