{ "METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002839","ANALYSIS_ID":"AN004646","VERSION":"1","CREATED_ON":"August 30, 2023, 4:35 am"}, "PROJECT":{"PROJECT_TITLE":"Metabolites alteration during ferroptotic process in tumor cells","PROJECT_SUMMARY":"Targeting ferroptosis, an iron-dependent form of regulated cell death triggered by the lethal overload of lipid peroxides, in cancer therapy is impeded by our limited understanding of the intersection of tumour’s metabolic feature and ferroptosis vulnerability. In the presented study, we performed metabolomics for cancer cell lines treated by ferroptosis inducer RSL3 or control DMSO.","INSTITUTE":"Zhongshan Hospital Fudan University","LAST_NAME":"Bi","FIRST_NAME":"Guoshu","ADDRESS":"Zhongshan Hospital, Fudan University, No. 180, Fenglin Road, Xuhui District, Shanghai, China","EMAIL":"18211210035@fudan.edu.cn","PHONE":"86 64041990"}, "STUDY":{"STUDY_TITLE":"Metabolic alteration during ferroptotic process in cancer cells.","STUDY_SUMMARY":"Targeting ferroptosis, an iron-dependent form of regulated cell death triggered by the lethal overload of lipid peroxides, in cancer therapy is impeded by our limited understanding of the intersection of tumour’s metabolic feature and ferroptosis vulnerability. In this study, we performed metabolomics in cancer cell lines pretreated with ferroptosis inducer RSL3 and control DMSO. We noted that the levels of a series of metabolites were significantly impacted by the RSL3 treatment, such as upregulated polyamines including putrescine, spermidine, and spermine. According to our previous study, we proved the pro-ferroptotic feature of polyamines in tumor cells, which was derived from the H2O2 produced during the polyamine metabolic process. This finding is consistent with our RNA-Seq data indicating upregulated ODC1 in the ferroptotic process. Therefore, it could be speculated that the RSL3-induced polyamine production leads to a positive-feedback loop that amplifies ferroptosis in tumor cells.","INSTITUTE":"Zhongshan Hospital Fudan University","LAST_NAME":"Bi","FIRST_NAME":"Guoshu","ADDRESS":"Zhongshan Hospital, Fudan University, No. 180, Fenglin Road, Xuhui District, Shanghai, China","EMAIL":"18211210035@fudan.edu.cn","PHONE":"86 64041990"}, "SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"}, "SUBJECT_SAMPLE_FACTORS":[ { "Subject ID":"-", "Sample ID":"ND1", "Factors":{"Treatment":"Control"}, "Additional sample data":{"genotype":"Wild-type","RAW_FILE_NAME":"ND1.raw"} }, { "Subject ID":"-", "Sample ID":"ND2", "Factors":{"Treatment":"Control"}, "Additional sample data":{"genotype":"Wild-type","RAW_FILE_NAME":"ND2.raw"} }, { "Subject ID":"-", "Sample ID":"NR1", "Factors":{"Treatment":"RSL3"}, "Additional sample data":{"genotype":"Wild-type","RAW_FILE_NAME":"NR1.raw"} }, { "Subject ID":"-", "Sample ID":"NR2", "Factors":{"Treatment":"RSL3"}, "Additional sample data":{"genotype":"Wild-type","RAW_FILE_NAME":"NR2.raw"} } ], "COLLECTION":{"COLLECTION_SUMMARY":"For cell samples, after being washed with ice-cold PBS, the cells were collected using a cell scraper and centrifuged for 5 min at 1,000 g. The samples were homogenized in liquid N2 for 1 min and defrosted in 4°C.","SAMPLE_TYPE":"Tumor cells"}, "TREATMENT":{"TREATMENT_SUMMARY":"A549 cells cultured in DMEM medium were treated with 0.5 uM RSL3 or DMSO for 24 h."}, "SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"After homogenization in liquid N2, 200 μL 80% methyl alcohol was added, followed by vortex for 1 min, sonication for 30 min at 4°C, and stand for 1 h at -20°C. Then, the samples were centrifuged at 12,000 rpm for 15 min at 4°C, and the supernatant was collected for subsequent LC-MS analysis."}, "CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"LC","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um)","SOLVENT_A":"Water containing 0.1% methanoic acid","SOLVENT_B":"Acetonitrile containing 0.1% methanoic acid","FLOW_GRADIENT":"Time:(0.01-2-4-8-10-14-15-15.1-16); Solvent A%: (95-95-70-50-20-0-0-95-95);Solvent B%: (5-5-30-50-80-100-100-5-5)","FLOW_RATE":"0.35 mL/min","COLUMN_TEMPERATURE":"45"}, "ANALYSIS":{"ANALYSIS_TYPE":"MS"}, "MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"Mass Scan Range 100-1200 Resolution (full scan) 70000 Resolution (HCD MS/MS scans) 17500 Spray Voltage (V) -3000 Sheath Gas Flow Rate (Arb) 35 Aux Gas Flow Rate (Arb) 8 Capillary Temperature (°C) 320","MS_RESULTS_FILE":"ST002839_AN004646_Results.txt UNITS:pmol/l Has m/z:Yes Has RT:Yes RT units:Minutes"} }