The early effects of cortisol on 3H-uridine and 14C-leucine uptake and incorporation in cell line I-407

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Charles Richard Lohrman (Creator)
The University of North Carolina at Greensboro (UNCG )
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Laura Anderton

Abstract: The purpose of this study was to investigate the effect of cortisol on uridine and leucine metabolism in cell line 1-407. Experiments were performed to determine the early effects of cortisol on the cells. Monolayered cell cultures were exposed to cortisol, labelled precursors such as 3H-uridine and 14C-leucine and metabolic inhibitors. These cells were then homogenized and subjected to trichloroacetic acid (TCA) separation. Radioactive precursors in the TCA-soluble and insoluble fractions acted as a measure of precursor uptake and incorporation. Preparatory experiments were performed to determine the fetal bovine serum (FBS) concentration for optimal cell growth. These experiments were also used to determine the cortisol concentration which produced the most consistent response. It was observed that cells grew best in 10% FBS. The lowest cortisol concentration which produced a consistently reproducible response in the cells was 0.05 ug/ml (1.7 x 10-7). This study demonstrated, for the first time, the early and continuous parallel hormonal response to uridine uptake and incorporation in an in vitro cell system. Cortisol stimulated an early (15 minute) increase in both uptake and incorporation of 3H-uridine followed by a rapid decrease at 30 minutes. The results of longer hormone incubations (2-4 hours) showed a decrease in specific activity of 3H-nucleoside in both TCA-soluble and TCA-insoluble fractions

Additional Information

Language: English
Date: 1977

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