Polyamine profiles and growth properties of ornithine decarboxylase overexpressing MCF-7 breast cancer cells in culture |
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Authors: | Andrea Manni Rita Wechter Rhea Grove Lisa Wei Julianne Martel Laurence Demers |
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Affiliation: | (1) Departments of Medicine, Pennsylvania State University College of Medicine, 500 University Drive, 17033 Hershey, PA, USA;(2) Pathology, Pennsylvania State University College of Medicine, 500 University Drive, 17033 Hershey, PA, USA;(3) Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, 500 University Drive, 17033 Hershey, PA, USA;(4) Center for Biostatistics and Epidemiology, Pennsylvania State University College of Medicine, 500 University Drive, 17033 Hershey, PA, USA;(5) Division of Endocrinology, Pennsylvania State University College of Medicine, 500 University Drive, P.O. Box 850, 17033 Hershey, PA, USA |
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Abstract: | Summary To determine the direct influence of the polyamine (PA) pathway on breast cancer phenotype, we employed a transfection approach to induce overexpression of the PA biosynthetic enzyme ornithine decarboxylase (ODC) in the hormone-responsive MCF-7 breast cancer cell line. Using a modified calcium phosphate method and an ODC cDNA coding for a truncated and more stable enzyme, we were able to achieve a moderate to marked degree of ODC overexpression (up to 150-fold) in a transient transfection system. ODC-overexpressing MCF-7 cells exhibited a selective increase in cellular putrescine content, while the levels of spermidine and spermine remained unaffected. Under defined culture conditions, overexpression of ODC resulted in a consistent but modest increase in [3H]thymidine incorporation into DNA which was similar in the presence and absence of 17- -estradiol, TGF- , and IGF-I. In the presence of serum, the effect of ODC overexpression on basal [3H]-thymidine incorporation into DNA was inconsistent, possibly as a result of subtle differences in culture conditions. Overall, our results support the hypothesis that activation of the PA biosynthetic pathway may confer a growth advantage to breast cancer cells. |
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Keywords: | breast cancer ornithine decarboxylase polyamines |
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