Human HGSOC Ovarian Cancer Metabolomics (Primary v. Omental Metastases)
Raw mass spectrometry data of matched primary ovarian tumors and omental metastases from human HGSOC samples (n=8 pairs). Metabolomics analysis was carried out by the Rutgers Metabolomics Core for manuscript entitled "G6PD Inhibition Sensitizes Ovarian Cancer Cells to Oxidative Stress in the Metastatic Omental Microenvironment". Services, results and/or products in support of the research project were generated by the Rutgers Cancer Institute of New Jersey Metabolomics Shared Resource, supported, in part, with funding from NCI-CCSG P30CA072720-5923.
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For human sample assays, tissue was ground using mortar and pestle while maintained in liquid nitrogen, and ~20 mg of tissue was used for DCFH-DA assays, G6PD enzyme activity assays, and WBs. For metabolomics analysis, metabolite extraction from ground tissue normalized by weight was performed in cold extraction buffer (40% methanol: 40% acetonitrile: 20% water) solution. Extraction was performed by vortexing and centrifugation at 16,000 x g for 10 min, after which the supernatant was isolated and 15% NH4HCO3 was added to neutralize acids and precipitate proteins. Following centrifugation, the supernatant was applied to a speed vac and a concentrated metabolite pellet was obtained. Following reconstitution, the sample was used for LC-MS performed at the Rutgers Metabolomics Core Service.