MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14072/m_MTBLS14072_LC-MS_positive_hilic_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14072/m_MTBLS14072_LC-MS_negative_hilic_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14072/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14072/a_MTBLS14072_LC-MS_positive_hilic_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14072/s_MTBLS14072.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14072/a_MTBLS14072_LC-MS_negative_hilic_metabolite_profiling.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14072ovarian cancer<p>The raw data collected using MassLynx V4.2 is processed by Progenesis QI software for peak extraction, peak alignment and other data processing operations, based on the Progenesis QI software online METLIN database and self-built library for identification.</p>mass spectrometry assay<p>Cell samples were transferred to an EP tube using 1000 μL of extraction solvent (methanol:acetonitrile = 1:1, containing 20 mg/L internal standard) in three aliquots (300 μL, 300 μL, and 400 μL), followed by vortexing for 30 seconds. Steel beads were added, and the samples were ground at 45 Hz for 10 minutes, then subjected to ultrasonication in an ice-water bath for 10 minutes. After incubation at –20°C for 1 hour, the samples were centrifuged at 12,000 rpm for 15 minutes at 4°C. A 500 μL aliquot of the supernatant was collected, dried using a vacuum concentrator, and reconstituted in 160 μL of acetonitrile:water (1:1). The mixture was vortexed for 30 seconds, sonicated in an ice-water bath for 10 minutes, and centrifuged again under the same conditions. Finally, 120 μL of the supernatant was transferred to an autosampler vial, and a 10 μL aliquot from each sample was pooled to prepare a QC sample for analysis.</p>Homo sapiens<p>After normalizing the original peak area information with the total peak area, the follow-up analysis was performed. Principal component analysis and Spearman correlation analysis were used to judge the repeatability of the samples within group and the quanlity control samples. The identified compounds are searched for classification and pathway information in KEGG, HMDB and lipidmaps databases.According to the grouping information, calculate and compare the difference multiples, T test was used to calculate the difference significance pvalue of each compound. The R language package ropls was used to perform OPLS-DA modeling, and 200 times permutation tests was performed to verify the reliability of the model. The VIP value of the model was calculated using multiple cross-validation. The method of combining the difference multiple, the P value and the VIP value of the OPLS-DA model was adopted to screen the differential metabolites. The screening criteria are FC&gt;1, P value&lt;0.05 and VIP&gt;1. The difference metabolites of KEGG pathway enrichment significance were calculated using hypergeometric distribution test.</p>Olaparibhttps://www.ebi.ac.uk/metabolights/MTBLS14072mahanlin@sdu.edu.cn<p>After removing the culture medium, wash the SK and SK/ola cells twice with ice-cold PBS and aspirate completely. Detach the cells using trypsin until they round up, then neutralize the trypsin by adding fresh medium. Remove the medium and gently resuspend the cells in PBS to obtain a single-cell suspension. Quickly count the cells and aliquot the volume corresponding to 1e7 cells. Immediately transfer this aliquot into 5 volumes of pre-chilled quenching reagent and vortex briefly for 10 seconds. Centrifuge at 1000g for 1 minute at 4°C in a pre-cooled rotor. Discard the supernatant, flash-freeze the cell pellet in liquid nitrogen for 30 seconds, and then store it at -80°C.</p><p><br></p><p><br></p><p><br></p>MetaboLightsPublicMetabolomicsLiquid Chromatography MS - positive - hilicLiquid Chromatography MS - negative - hilicolaparib-resistant cellsWaters ACQUITY UPLC H-Class Systemovarian canceruntargeted analysisuntargeted metabolitesHomo sapiensWaters UPLC Xevo G2-XS QTOFcell<p>The LC/MS system for metabolomics analysis is composed of Waters Acquity I-Class PLUS ultra-high performance liquid tandem Waters Xevo G2-XS QTof high resolution mass spectrometer. The column used is purchased from Waters Acquity UPLC HSS T3 column (1.8um 2.1*100mm)&nbsp;</p><p>Positive ion mode: mobile phase A: 0.1% formic acid aqueous solution; mobile phase B: 0.1% formic acid acetonitrile&nbsp;</p><p>Negative ion mode: mobile phase A: 0.1% formic acid aqueous solution; mobile phase B: 0.1% formic acid acetonitrile&nbsp;</p><p>Injection volume 2μL</p><p>&nbsp;</p>Targeting SLC1A5-mediated glutamine metabolism overcomes PARP inhibitor resistance in ovarian cancer.olaparib-resistant cellsWaters ACQUITY UPLC H-Class Systemovarian canceruntargeted analysisuntargeted metabolitesHomo sapiensWaters UPLC Xevo G2-XS QTOFcellHanlin MaShandong University<p>Waters Xevo G2-XS QTOF high resolution mass spectrometer can collect primary and secondary mass spectrometry data in MSe mode under the control of the acquisition software (MassLynx V4.2, Waters). In each data acquisition cycle, dual-channel data acquisition can be performed on both low collision energy and high collision energy at the same time. The low collision energy is off, the high collision energy range is 10~40V, and the scanning frequency is 0.2 seconds for a mass spectrum. The parameters of the ESI ion source are as follows: Capillary voltage: 2500V (positive ion mode) or -2000V (negative ion mode); cone voltage: 30V; ion source temperature: 100°C; desolvent gas temperature 500°C; backflush gas flow rate: 50L/ h; Desolventizing gas flow rate: 800L/h.</p>falseTargeting SLC1A5-mediated glutamine metabolism overcomes PARP inhibitor resistance in ovarian cancer<p>Poly(ADP-ribose) polymerase inhibitors (PARPi), such as olaparib, have significantly improved outcomes in ovarian cancer (OC); however, therapy resistance remains a major clinical challenge. Here, we showed that glutamine metabolism was upregulated in PARPi-resistant OC cells and identified the glutamine transporter SLC1A5 as a key mediator of this metabolic adaptation. Inhibition of SLC1A5 increased the sensitivity of OC cells to olaparib both in vitro and in vivo. Mechanistically, we demonstrated that the deubiquitinating enzyme USP14 directly interacted with and stabilized SLC1A5 by removing K48-linked polyubiquitin chains, thereby enhancing glutamine uptake, promoting glutathione synthesis, alleviating oxidative stress, and activating the mTORC1 pathway. This metabolic rewiring improved DNA damage repair capacity and ultimately contributed to PARPi resistance. Notably, combined treatment with the SLC1A5 inhibitor V-9302 and olaparib synergistically suppressed tumor growth in a patient-derived xenograft (PDX) model. Collectively, our findings identify the USP14–SLC1A5 axis as a novel regulator of PARPi resistance and suggest that targeting glutamine metabolism represents a promising therapeutic strategy to overcome PARPi resistance in ovarian cancer.</p>2026-04-082026-03-18MTBLS14072