Abstract
Abstract Stable isotope-resolved metabolomics (SIRM) provides a comprehensive and functional readout of cellular activity and is therefore a useful tool both for biomarker discovery and fundamental cancer biology. We have established a primary lung squamous cell carcinoma line (PSLC1), isolated from the lung tumor tissue of a non-small cell lung cancer patient, for which the metabolic profile has been established. We used SIRM to characterize the metabolic activity of the cells in mouse xenografts and in cell culture. Here we report the metabolic activities of the cell line in culture using two different tracers to evaluate central metabolism. PSLC1 cells were grown in either [U-13C]-glucose or [U-13C,15N]-glutamine labeled medium. Cell extracts and medium were prepared and analyzed by 1H-NMR, HSQC, TOCSY, GC-MS, and FTICR-MS. Cells exhibited linear glucose consumption that persisted even as proliferation plateaued. The conversion of glucose to lactate occurred at a strikingly high 89%. Anaplerotic pathways involving pyruvate carboxylation and glutaminolysis were active and appeared to be dependent on cellular proliferation. Glutamine represented an important contributor for biosynthetic and bioenergetic cellular needs, including de novo pyrimidine biosynthesis and glucose-independent TCA cycling. In addition, glutamine served as the major precursor in proline and glutathione biosynthesis. Distinct glutamine utilization and labeling patterns indicated that this cancer might be driven by C-MYC overexpression. C-MYC overexpression was observed by targeted protein analysis guided by metabolomics-edited transcriptomics analysis (Pro-META). The ability of SIRM to generate hypotheses about protein expression demonstrates its potential use as a first line biomarker detection method. Comparison with upstream in situ and xenograft data will allow us to assess model validity and the effects of tumor microenvironment. In turn, the in vitro data will help us interpret human in situ data more robustly. Supported by grant R25-CA-134283 and 1P01CA163223-01A1 from the National Cancer Institute Citation Format: Connor J. Kinslow, Teresa W-M Fan, Jin L. Tan, Pawel K. Lorkiewicz, Ramya Balasubramaniam, Yihua Cai, Jun Yan, Andrew N. Lane. Validation of primary non-small cell lung cancer model: Stable isotope-resolved metabolomics (SIRM) reveals functional biochemistry. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1422. doi:10.1158/1538-7445.AM2014-1422
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