Stevens, Axel P. and Dettmer, Katja and Wallner, Susanne and Bosserhoff, Anja K. and Oefner, Peter J. (2008) Quantitative analysis of 5 '-deoxy-5 '-methylthioadenosine in melanoma cells by liquid chromatography-stable isotope ratio tandem mass spectrometry. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 876 (1). pp. 123-128. ISSN 1570-0232, 1873-376X
Full text not available from this repository. (Request a copy)Abstract
The frequent deletion of the human chromosomal region 9p21, including the methylthioadenosine phosphorylase (MTAP) gene, is hypothesized to lead to the intra- and/or extracellular accumulation of 5'-(deoxy-5'-methylthioadenosine(MTA) in cancer cells and the subsequent promotion of tumor progression. The lack of sensitive methodology for the direct measurement of MTA in tumor cells has hampered the testing of this hypothesis to date. A liquid chromatography electrospray ionization tandem mass spectrometry method (LC-MS/MS) was developed for the absolute quantitative determination of MTA in cell culture media and cell extracts using stable isotope labeled MTA as an internal standard. Limit of detection (LOD) and lower limit Of quantification (LLOQ) were 62.5 pM and 2 W respectively, and allowed the direct measurement of MTA in biological samples without prior enrichment. Average imprecision of MTA extraction from cells and cell media, as well as LC-MS/MS analysis were 9.7, 3.8 and 1.9%, respectively. The method enabled the demonstration of the accumulation of MTA in melanoma cell culture media reaching a steady-state level within 24 h. Only a slight difference in extracellular MTA concentrations was observed between cells with and without MTAP expression. However, there was a fourfold increase in intracellular MTA concentration in melanoma cells lacking MTAP, thus confirming the hypothesized accumulation of MTA in human cancer cells harboring a chromosome 9p21 deletion. (C) 2008 Elsevier B.V. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PURINE METABOLIC ENZYME; METHYLTHIOADENOSINE PHOSPHORYLASE; POLYAMINE METABOLISM; URINARY NUCLEOSIDES; MALIGNANT-MELANOMA; 5-DEOXY-5-METHYLTHIOADENOSINE; DEFICIENT; LINES; ACID; 5 '-Deoxy-5 '-methylthioadenosine; Methylthioadenosine phosphorylase; LC-MS/MS; Stable isotope dilution; Melanoma |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Institut für Funktionelle Genomik > Lehrstuhl für Funktionelle Genomik (Prof. Oefner) Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Oct 2020 12:29 |
| Last Modified: | 14 Oct 2020 12:29 |
| URI: | https://pred.uni-regensburg.de/id/eprint/29972 |
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