The metabolites were identified by searching database, and the main databases were the HMDB (http://www.hmdb.ca/), Metlin ( https://metlin.scripps.edu/) and the self-compiled Majorbio Database (MJDB) of Majorbio Biotechnology Co., Ltd. (Shanghai, China).
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative - reverse-phase","Liquid Chromatography MS - positive - reverse-phase"],"chromatography_protocol":["Chromatographic separation was performed on a Thermo Scientific Vanquish Horizon UHPLC system coupled to a Thermo Scientific Q Exactive HF-X mass spectrometer. Metabolites were separated using an ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm internal diameter, 1.8 μm particle size; Waters, Milford, MA, USA), maintained at 40 °C. The injection volume was 3 μL .
Mobile phase A consisted of water:acetonitrile (95:5, v/v) containing 0.1% formic acid. Mobile phase B consisted of acetonitrile:isopropanol:water (47.5:47.5:5, v/v/v) containing 0.1% formic acid.
For electrospray negative-ion mode, the gradient programme was as follows: 0.0 min, 100% A and 0% B; 1.5 min, 95% A and 5% B; 2.0 min, 90% A and 10% B; 4.5 min, 70% A and 30% B; 5.0 min, 0% A and 100% B; 6.3 min, 0% A and 100% B; 6.4 min, 100% A and 0% B; and 8.0 min, 100% A and 0% B. The flow rate was maintained at 0.4 mL/min throughout the run.
"],"publication":["JNK Inhibition Overcomes DHODH Resistance in KRAS-Mutant Cancer by Converting Metabolic Stasis into Dual-Gasdermin Lytic Cell Death."],"submitter_name":["Pan Li"],"submitter_affiliation":["Xuzhou Medicine University"],"organism_part":["HCT-116 cell"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["Cell pellets were placed in 2 mL centrifuge tubes. One 6-mm grinding bead and 400 μL of methanol:water (4:1, v/v) extraction solvent containing four internal standards, including L-2-chlorophenylalanine (0.02 mg/mL), were added to each sample.
Samples were homogenized using a cryogenic tissue grinder for 6 min at −10 °C and 50 Hz. Metabolites were then extracted by ultrasonication for 30 min at 5 °C and 40 kHz. Samples were incubated at −20 °C for 30 min and centrifuged at 13,000 × g for 15 min at 4 °C. The resulting supernatants were transferred to autosampler vials with inserts for LC-MS analysis.
A pooled quality-control (QC) sample was prepared by combining 20 μL of supernatant from each study sample.
"],"organism":["Homo sapiens"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS15162"],"author":["Sun Yang. Xuzhou Medical University. yangsun@xzhmu.edu.cn.","Pan Li. Xuzhou Medical University. panli@xzhmu.edu.cn."],"data_transformation_protocol":["The pretreatment of LC/MS raw data was performed by Progenesis QI (Waters Corporation, Milford, USA) software, and a three-dimensional data matrix in CSV format was exported. The information in this three-dimensional matrix included: sample information, metabolite name and mass spectral response intensity. Internal standard peaks, as well as any known false positive peaks (including noise, column bleed, and derivatized reagent peaks), were removed from the data matrix, deredundant and peak pooled.
"],"study_factor":["Experimental group"],"submitter_email":["panli@xzhmu.edu.cn"],"sample_collection_protocol":["Human HCT116 colorectal carcinoma cells were used for untargeted metabolomics analysis. Cells were assigned to four treatment groups and treated for 24 h: (1) vehicle control, treated with DMSO; (2) Brequinar (BRQ), treated with 1 μM BRQ; (3) JNK-IN-8, treated with 1 μM JNK-IN-8; and (4) combined treatment, treated with 1 μM BRQ plus 1 μM JNK-IN-8. Each group contained six biological replicates (n = 6), giving 24 study samples in total. Approximately 1.5 × 10^7 cells were collected for each replicate. Cell pellets were stored at −80 °C until metabolite extraction.
"],"omics_type":["Metabolomics"],"study_design":["ultra-performance liquid chromatography-mass spectrometry","Metabolomics Platform, Shanghai Majorbio Bio-pharm Technology Co., Ltd., Shanghai, China","Metabolomics","Thermo Scientific Vanquish Horizon UHPLC system","colorectal cancer","Q Exactive HF-X","not applicable","untargeted analysis","Thermo Scientific Q Exactive HF-X","Homo sapiens","cell","Progenesis QI"],"curator_keywords":["ultra-performance liquid chromatography-mass spectrometry","Metabolomics Platform, Shanghai Majorbio Bio-pharm Technology Co., Ltd., Shanghai, China","Metabolomics","Thermo Scientific Vanquish Horizon UHPLC system","colorectal cancer","not applicable","Q Exactive HF-X","untargeted analysis","Thermo Scientific Q Exactive HF-X","Homo sapiens","cell","Progenesis QI"],"mass_spectrometry_protocol":["Mass spectrometric analysis was performed using a Thermo Scientific Q Exactive HF-X high-resolution mass spectrometer equipped with an electrospray ionization (ESI) source. Data were acquired in electrospray negative-ion mode.
The ESI source parameters in negative-ion mode were as follows: spray voltage, −3.5 kV; sheath gas flow rate, 30 arbitrary units; auxiliary gas flow rate, 25 arbitrary units; capillary temperature, 350 °C; and S-lens RF level, 50.
Full-scan MS spectra were acquired over an m/z range of 70–1,050 at a resolution of 60,000. MS/MS spectra were acquired at a resolution of 7,500 using stepped normalized collision energies of 20, 40, and 60 eV. MS and MS/MS spectral information was used for untargeted metabolite profiling and metabolite annotation.Please update this protocol description
"],"additional_accession":[]},"is_claimable":false,"name":"Untargeted metabolomic profiling of HCT116 cells following DHODH and JNK inhibition","description":"Untargeted LC-MS metabolomics was used to investigate metabolic alterations associated with DHODH inhibition and JNK signaling in colorectal cancer. The study included control, DHODH inhibitor treated, JNK inhibitor treated, and combined treatment groups, with pooled quality control samples for analytical stability assessment.
","dates":{"publication":"2026-08-13","submission":"2026-07-27"},"accession":"MTBLS15162","cross_references":{}}