MetaboLights

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identification of metabolites was performed by comparing of accuracy m/z value (<10 ppm), and MS/MS spectra with an in-house database established with available authentic standards.MetaboLightsPublicLiquid Chromatography MS - positive - hilicLiquid Chromatography MS - negative - hilicAnalysis was performed using an UHPLC (1290 Infinity LC, Agilent Technologies) coupled to a quadrupole time-of-flight (AB Sciex TripleTOF 6600) in Shanghai Applied Protein Technology Co., Ltd. For HILIC separation, samples were analyzed using a 2.1 mm x 100 mm ACQUIY UPLC BEH Amide 1.7 µm column (waters, Ireland). In both ESI positive and negative modes, the mobile phase contained A = 25 mM ammonium acetate and 25 mM ammonium hydroxide in water and B = acetonitrile. The gradient was 95% B for 0.5 min and was linearly reduced to 65% in 6.5 min, and then was reduced to 40% in 1.0 min and kept for 1.0 min, and then increased to 95% in 0.1 min, with a 3.0 min re-equilibration period employed.Choline metabolism drives metastasis in BRCA1-deficient ovarian cancers by activating FAM3C.China Pharmaceutical UniversityJiawei ZhuCellsmass spectrometry assayFor LC-MS analysis, the samples were re-dissolved in 100 μL acetonitrile/water (1:1, v/v) solvent and centrifuged at 14,000 x g at 4 °C for 15 min, then the supernatant was injected. To monitor the stability and repeatability of instrument analysis, quality control (QC) samples were prepared by pooling 10 μL of each sample and analyzed together with the other samples. The QC samples were inserted regularly and analyzed in every 5 samples.Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS12245Yue Wu. triplewu@foxmail.com.Jiawei Zhu. 3124124442@stu.cpu.edu.cn.Wanying Zhang. zhangwanying178@163.com.Cong Wang. 19850855806@163.com.Guo Chen. China Pharmaceutical University. 639 Longmian Avenue, Nanjing, Jiangsu 211198, China. gchen84@cpu.edu.cn.Xuexia Xie. 1425765149@qq.com.Dake Li. lidake2002@163.com.Jiahuang Li. lijiah@cpu.edu.cn.Binghe Sun. 18560380238@163.com.The raw MS data were converted to mzXML files using ProteoWizard MSConvert before importing into freely available XCMS software. For peak picking, the following parameters were used: centWave, m/z = 10 ppm, peakwidth = c(10,60), prefilter = c(10,100). For peak grouping, bw = 5, mzwid = 0.025, minfrac = 0.5 were used. CAMERA (Collection of Algorithms of MEtabolite pRofile Annotation) was sued for annotation of isotopes and adducts. In the extracted ion features, only the variables having more than 50% of the nonzero measurement values in at least one group were kept. Compound identification of metabolites was performed by comparing of accuracy m/z value (<10 ppm), and MS/MS spectra with an in-house database established with available authentic standards. After sum-normalization, the processed data were analyzed by R package (ropls), where it was subjected to multivariate data analysis, including Pareto-scaled principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA).Treatment3124124442@stu.cpu.edu.cnThe culture medium from the cultured Hela cells (~107 cells per sample) was removed using pipette. Then the cells were washed with PBS under 37 °C and the PBS was removed. Then 800 μL of cold methanol/acetonitrile (1:1, v/v) to remove the protein and extract the metabolites. The mixture was collected into a new centrifuge tube, and centrifuged at 14,000 x g for 20 min to collect the supernatant. The supernatant was dried in a vacuum centrifuge.Metabolomicsultra-performance liquid chromatography-mass spectrometrypooled quality control samplecancer cellAgilent Technologies 1290 Infinity LCuntargeted analysisHomo sapiensuntargeted metabolitesAB SCIEX TripleTOF 6600Cellsexperimental sampleultra-performance liquid chromatography-mass spectrometrypooled quality control samplecancer cellAgilent Technologies 1290 Infinity LCuntargeted analysisHomo sapiensuntargeted metabolitesAB SCIEX TripleTOF 6600Cellsexperimental sampleThe ESI source conditions were set as follows: Ion Source Gas1 (Gas1) as 60, Ion Source Gas2 (Gas2) as 60, curtain gas (CUR) as 30, source temperature as 600 °C, IonSpray Voltage Floating (ISVF) ± 5500 V. In MS only acquisition, the instrument was set to acquire over the m/z range 60-1000 Da, and the accumulation time for TOF MS scan was set at 0.20 s/spectra. In auto MS/MS acquisition, the instrument was set to acquire over the m/z range 25-1000 Da, and the accumulation time for product ion scan was set at 0.05 s/spectra. The product ion scan is acquired using information dependent acquisition (IDA) with high sensitivity mode selected. The parameters were set as follows: the collision energy (CE) was fixed at 35 V with ± 15 eV; declustering potential (DP), 60 V (+) and -60 V (-); exclude isotopes within 4 Da, candidate ions to monitor per cycle: 10.16-Hydroxypalmitic acidAllantoinCitrateErucic acidGuanosine diphosphate mannosealpha-D-Glucose 1-phosphatePalmitic acidPentadecanoic AcidChenodeoxycholateUndecanoic AcidDeoxyinosineD-Ribose 5-phosphateL-AlanineMaleic acid2'-Deoxyuridine4-Hydroxycinnamic acidStearic acidPyridoxamine 5'-phosphateUridine diphosphate glucose(UDP-D-Glucose)2'-Deoxyadenosine 5'-triphosphate (dATP)cis-AconitateL-Malic acidL-ProlineCapric acidMyristic acidDihydrouracil5-L-Glutamyl-L-alanineOxypurinol2-Oxoadipic acidD-ThreitolDodecanoic acidcis-4-Hydroxy-D-prolineFolateTridecanoic acid (Tridecylic acid)3-Methylphenylacetic acidOleic acidmyo-Inositol2'-Deoxycytidine diphosphate (dCDP)D-Ribose 1,5-bisphosphateDL-lactateHydroxyphenyllactic acidN-Acetyl-L-glutamateXanthosinePelargonic acidDeoxyguanosine triphosphate (dGTP)Linoleic acidN-Acetylglucosamine 1-phosphateD-Mannose 1-phosphateketoisocaproic acidCitraconic acidD-Erythrose 4-phosphateAdenosine 3',5'-diphosphate (PAP)falseMetabolic study of BRCA1 knockout cells and WT cells in homo sapiensBRCA1 mutations are a hallmark of hereditary ovarian cancer, strongly linked to deficiencies in homologous recombination (HR) DNA repair and impaired DNA replication fork protection. However, its roles in cancer progression beyond maintaining genomic integrity remain poorly understood. Through metabolomics approaches, we found BRCA1-deficiency strikingly increased choline metabolism. Loss of BRCA1 promotes choline uptake through upregulating choline transporter-like protein 4 (CTL4). BRCA1 directly binds and recruits EZH2-mediated H3K27Me3 deposition to CTL4 promoter. CTL4 was therefore overexpressed in ovarian cancer tissues with BRCA1 mutations. Furthermore, BRCA1-deficiency significantly promotes ovarian cancer invasion, while inhibition of CTL4 reverses the high metastatic potential of BRCA1-deficient ovarian cancer cells, suggesting the functionality and specificity of CTL4 as a therapeutic target. Additionally, we discovered that phosphocholine, the choline metabolite increased by CTL4 overexpression, interacted with and stabilized the epithelial-to-mesenchymal transition inducer FAM3C in BRCA1-deficient ovarian cancer cells. Importantly, we identified a potent CTL4 inhibitor, DT-13, which significantly reduces choline metabolism and effectively suppresses metastasis in BRCA1-deficient ovarian cancers. Therefore, our study uncovers a mechanism underlying metastasis in BRCA1-deficient cancers and identifies CTL4 as a therapeutic target for metastatic ovarian cancer patients with BRCA1 mutations.2026-05-152025-02-21MTBLS12245HMDB0036458HMDB0001563HMDB0000001/HMDB0000479HMDB0000699HMDB0007098/HMDB0007099/HMDB0007100/HMDB0007101/HMDB0007103/HMDB0007108/HMDB0007112/HMDB0007131/HMDB0007132/HMDB0007156/HMDB0007166/HMDB0007170/HMDB0007223/HMDB0007224/HMDB0007248/HMDB0007257/HMDB0007398HMDB0000564/HMDB0007878/HMDB0007879/HMDB0007880/HMDB0007882/HMDB0007884/HMDB0007886/HMDB0007887/HMDB0007897/HMDB0007898/HMDB0007911/HMDB0007912/HMDB0007913/HMDB0007914/HMDB0007936/HMDB0007937/HMDB0007938/HMDB0007939/HMDB0007940/HMDB0007958/HMDB0007959/HMDB0007960/HMDB0007961/HMDB0007964/HMDB0007966/HMDB0007967/HMDB0007981/HMDB0007982/HMDB0007983/HMDB0007984/HMDB0007985/HMDB0008002/HMDB0008003/HMDB0008004/HMDB0008005/HMDB0008028/HMDB0008029/HMDB0008065/HMDB0008066/HMDB0008067/HMDB0008068/HMDB0008069/HMDB0008070/HMDB0008071/HMDB0008091/HMDB0008092/HMDB0008119/HMDB0008120/HMDB0008121/HMDB0008126/HMDB0008129/HMDB0008145/HMDB0008147/HMDB0008182/HMDB0008183/HMDB0008186/HMDB0008191/HMDB0008192/HMDB0008230/HMDB0008231/HMDB0008232/HMDB0008233/HMDB0008269/HMDB0008270/HMDB0008271/HMDB0008272/HMDB0008274/HMDB0008304/HMDB0008306/HMDB0008308/HMDB0008309/HMDB0008312/HMDB0008317/HMDB0008322/HMDB0008323/HMDB0008324/HMDB0008375/HMDB0008377/HMDB0008379/HMDB0008400/HMDB0008401/HMDB0008402/HMDB0008409/HMDB0008410/HMDB0008413/HMDB0008414/HMDB0008415/HMDB0008416/HMDB0008426/HMDB0008427/HMDB0008428/HMDB0008429/HMDB0008431/HMDB0008444/HMDB0008507/HMDB0008508/HMDB0008509/HMDB0008510/HMDB0008514/HMDB0008515/HMDB0008516/HMDB0008522/HMDB0008536/HMDB0008537/HMDB0008538/HMDB0008539/HMDB0008540/HMDB0008541/HMDB0008542/HMDB0008543/HMDB0008606/HMDB0008607/HMDB0008608/HMDB0008609/HMDB0008610/HMDB0008611/HMDB0008612/HMDB0008614/HMDB0008615/HMDB0008655/HMDB0008657/HMDB0008658/HMDB0008659/HMDB0008660/HMDB0008661/HMDB0008662/HMDB0008663/HMDB0008664/HMDB0008665/HMDB0008668/HMDB0008669/HMDB0008670/HMDB0008671/HMDB0008672/HMDB0008673/HMDB0008674/HMDB0008676/HMDB0008681/HMDB0008682/HMDB0008683/HMDB0008684/HMDB0008686/HMDB0008688/HMDB0008690/HMDB0008691/HMDB0008692/HMDB0008697/HMDB0008698/HMDB0008699/HMDB0008700/HMDB0008702/HMDB0008703/HMDB0008704/HMDB0008706/HMDB0008707/HMDB0008730/HMDB0008731/HMDB0008732/HMDB0008733/HMDB0008734/HMDB0008735/HMDB0008736/HMDB0008737/HMDB0008738/HMDB0008745/HMDB0008746/HMDB0008747/HMDB0008748/HMDB0008749/HMDB0008750/HMDB0008751/HMDB0008752/HMDB0008754/HMDB0008758/HMDB0008761/HMDB0008762/HMDB0008766/HMDB0008768/HMDB0008770/HMDB0008773/HMDB0008774/HMDB0008775/HMDB0008791/HMDB0008792/HMDB0008793/HMDB0008795/HMDB0008796/HMDB0008798/HMDB0008799/HMDB0008800HMDB0000960HMDB0001044HMDB0031213HMDB0000403HMDB0002203HMDB0000387HMDB0003332HMDB0006023HMDB0000982HMDB0000280HMDB0000201HMDB0000895HMDB0001206HMDB0000034HMDB0000050HMDB0011616HMDB0000058HMDB0003540HMDB0001341HMDB0001003HMDB0000538HMDB0000045HMDB0001178HMDB0000481HMDB0003514HMDB0000052HMDB0000043/HMDB0059606HMDB0000229HMDB0001564HMDB0000097HMDB0000064HMDB0000562HMDB0031404HMDB0000089HMDB0001546HMDB0001413HMDB0001176HMDB0000082HMDB0000630HMDB0006483HMDB0000101HMDB0001342HMDB0014849HMDB0000779/HMDB0014712HMDB0000169HMDB0001078HMDB0003411HMDB0015109HMDB0001248HMDB0011737HMDB0003351HMDB0001095/HMDB0062554HMDB0000125HMDB0003337HMDB0000086HMDB0000133HMDB0001201HMDB0001397HMDB0001273HMDB0000725/HMDB0036576HMDB0000157HMDB0060484HMDB0000195HMDB0000646HMDB0000517HMDB0000168HMDB0000191HMDB0000033HMDB0000904HMDB0000574HMDB0000148HMDB0000177HMDB0000172HMDB0000684HMDB0000687HMDB0000182HMDB0000696HMDB0000159HMDB0000267/HMDB0000805/HMDB0060262HMDB0000167HMDB0000929HMDB0000158HMDB0000883HMDB0001262HMDB0060486HMDB0002000HMDB0003357HMDB0004044HMDB0002038HMDB0001325HMDB0001067HMDB0001062HMDB0000766HMDB0000812HMDB0000866HMDB0001015HMDB0001406HMDB0000902HMDB0000217HMDB0001488HMDB0001132/HMDB0059646HMDB0000224HMDB0000210HMDB0015563HMDB0000821HMDB0001511HMDB0001565HMDB0001545HMDB0000239HMDB0035837HMDB0001487HMDB0000244HMDB0001185HMDB0001173HMDB0000114HMDB0001348/HMDB0010168/HMDB0010169/HMDB0011698/HMDB0011699/HMDB0012084/HMDB0012085/HMDB0012087/HMDB0012088/HMDB0012089/HMDB0012090/HMDB0012091/HMDB0012092/HMDB0012093/HMDB0012094/HMDB0012095/HMDB0012096/HMDB0012100/HMDB0012101/HMDB0012102/HMDB0012103/HMDB0012104/HMDB0012105/HMDB0012107/HMDB0013461/HMDB0013462/HMDB0013463/HMDB0013464/HMDB0013465/HMDB0013466/HMDB0013467/HMDB0013468/HMDB0013469HMDB0000252HMDB0003553HMDB0000254HMDB0000251HMDB0001372HMDB0000273HMDB0000262HMDB0012204HMDB0032538HMDB0000306HMDB0000302/HMDB0062573HMDB0000290HMDB0000300HMDB0000296HMDB0000295HMDB0000935HMDB0000288HMDB0000285HMDB0000301/HMDB0034174HMDB0006294HMDB0001532HMDB0001245HMDB0000012HMDB0000225HMDB0002222HMDB0002035HMDB0006248HMDB0000061HMDB0000462HMDB0001586HMDB0000511HMDB0000518HMDB0060460HMDB0000072HMDB0000634HMDB0000094HMDB0001440HMDB0000071HMDB0001321HMDB0000076HMDB0001311/HMDB0000190HMDB0006330HMDB0000638HMDB0011688HMDB0001548HMDB0004136HMDB0002068HMDB0000121HMDB0001163/HMDB0062759HMDB0000755HMDB0000695HMDB0000161HMDB0000673HMDB0000156HMDB0000162HMDB0000176HMDB0000211HMD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