#METABOLOMICS WORKBENCH amandanpope_20250708_132206 DATATRACK_ID:6147 STUDY_ID:ST004427 ANALYSIS_ID:AN007406 PROJECT_ID:PR002798 VERSION 1 CREATED_ON December 9, 2025, 1:59 pm #PROJECT PR:PROJECT_TITLE Brief Pulses of High-Level Fluid Shear Stress Enhance Metastatic Potential and PR:PROJECT_TITLE Rapidly Alter the Metabolism of Cancer Cells PR:PROJECT_TYPE GC-and LC-MS PR:PROJECT_SUMMARY Circulating tumor cells (CTCs) face challenges to their survival including PR:PROJECT_SUMMARY mechanical and oxidative stresses that are different from cancer cells in solid PR:PROJECT_SUMMARY primary and metastatic tumors. The impact of adaptations to the fluid PR:PROJECT_SUMMARY microenvironment of the circulation on the outcome of the metastatic cascade are PR:PROJECT_SUMMARY not well understood. Here we find that cancer cells (PC-3, MDA-MB-231, Myc-CaP) PR:PROJECT_SUMMARY exposed to brief pulses of high-level FSS exhibit enhanced invasiveness and PR:PROJECT_SUMMARY anchorage-independent proliferation in vitro and enhanced metastatic PR:PROJECT_SUMMARY colonization/tumor formation in vivo. Cancer cells exposed to FSS rapidly alter PR:PROJECT_SUMMARY their metabolism in a manner that promotes survival by providing energy for PR:PROJECT_SUMMARY cytoskeletal remodeling and contractility as well as reducing equivalents to PR:PROJECT_SUMMARY counter oxidative stress associated with cell detachment. Thus, exposure to FSS PR:PROJECT_SUMMARY may provide CTCs an unexpected survival benefit that promotes metastatic PR:PROJECT_SUMMARY colonization. PR:INSTITUTE Eastern Virginia Medical School at Old Dominion University PR:DEPARTMENT Department of Biomedical & Translational Sciences PR:LABORATORY Dr. Michael Henry PR:LAST_NAME Henry PR:FIRST_NAME Michael PR:ADDRESS 700 W Olney Rd, PO Box 1980, Norfolk, VA, 23501 PR:EMAIL mhenry@odu.edu PR:PHONE 757-446-5619 PR:FUNDING_SOURCE NIH PR:PUBLICATIONS doi: https://doi.org/10.1101/2025.04.09.647247 PR:CONTRIBUTORS Amanda N Pope, Devon L Moose, Eric B Taylor #STUDY ST:STUDY_TITLE Brief Pulses of High-Level Fluid Shear Stress Enhance Metastatic Potential and ST:STUDY_TITLE Rapidly Alter the Metabolism of Cancer Cells ST:STUDY_TYPE MS studies on PC-3 human prostate cancer and MDA-MB-231 human breast cancer cell ST:STUDY_TYPE lines ST:STUDY_SUMMARY PC-3 and MDA-MB-231 cell lines were grown in complete media at 37C until 80% ST:STUDY_SUMMARY confluency. The cells were either held in suspension as static controls or ST:STUDY_SUMMARY exposed to fluid shear stress (FSS) as outlined in PMID:32187555. Once the final ST:STUDY_SUMMARY pulse of FSS was conducted the cells were then plunged into ice cold PBS, ST:STUDY_SUMMARY counted, centrifuged, and cell pellet flash frozen in liquid nitrogen. Cell ST:STUDY_SUMMARY pellets were stored at -80C until profiling was initiated. Three biological ST:STUDY_SUMMARY replicates and match controls were submitted for metabolomic profiling. The data ST:STUDY_SUMMARY from the metabolomic profiling revealed that immediately upon FSS exposure, ST:STUDY_SUMMARY cancer cells significantly utilize ATP to support the energetically costly ST:STUDY_SUMMARY events of cytoskeleton rearrangement. We additionally observed significant ST:STUDY_SUMMARY utilization in amino acids including serine, threonine, and methionine-- which ST:STUDY_SUMMARY suggests utilization of the one-carbon cycle. We identified a slight increase of ST:STUDY_SUMMARY lactate production in the PC-3 cells, which was not observed in the MDA-MB-231 ST:STUDY_SUMMARY cells. This suggests a potential difference in energy production methods between ST:STUDY_SUMMARY the two cancer types during FSS exposure. Herein, this metabolomic profile ST:STUDY_SUMMARY demonstrates immediate activation of metabolic pathways to support the energy ST:STUDY_SUMMARY demand that is required to resist destruction to FSS. ST:INSTITUTE Eastern Virginia Medical School at Old Dominion University ST:DEPARTMENT Department of Biomedical & Translational Sciences ST:LAST_NAME Henry ST:FIRST_NAME Michael ST:ADDRESS 700 W Olney Rd, PO Box 1980, Norfolk, VA, 23501 ST:EMAIL mhenry@odu.edu ST:PHONE 757-446-5619 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 24 ST:PUBLICATIONS Pope, AN; Moose, DL, et al. Brief Pulses of High-Level Fluid Shear Stress ST:PUBLICATIONS Enhance Metastatic Potential and Rapidly Alter the Metabolism of Cancer Cells. ST:PUBLICATIONS bioRxiv 2025 , doi: https://doi.org/10.1101/2025.04.09.647247 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Male and female SU:CELL_BIOSOURCE_OR_SUPPLIER ATCC SU:CELL_PASSAGE_NUMBER P5 SU:CELL_COUNTS 2E6 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS LC-MS MDA_U1_LC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=S02_01029_MH_AP_20240416_QE1_19.raw SUBJECT_SAMPLE_FACTORS LC-MS MDA_U2_LC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=S06_01029_MH_AP_20240416_QE1_16.raw SUBJECT_SAMPLE_FACTORS LC-MS MDA_U3_LC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=S10_01029_MH_AP_20240416_QE1_23.raw SUBJECT_SAMPLE_FACTORS LC-MS MDA_U4_LC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=S14_01029_MH_AP_20240416_QE1_24.raw SUBJECT_SAMPLE_FACTORS LC-MS MDA_S1_LC Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=S04_01029_MH_AP_20240416_QE1_8.raw SUBJECT_SAMPLE_FACTORS LC-MS MDA_S2 Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=S08_01029_MH_AP_20240416_QE1_22.raw SUBJECT_SAMPLE_FACTORS LC-MS MDA_S3_LC Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=S12_01029_MH_AP_20240416_QE1_9.raw SUBJECT_SAMPLE_FACTORS LC-MS MDA_S4_LC Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=S16_01029_MH_AP_20240416_QE1_20.raw SUBJECT_SAMPLE_FACTORS blank QC1_MDA_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC1_01029_MH_AP_20240416_QE1_5.raw SUBJECT_SAMPLE_FACTORS blank QC2_MDA_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC2_01029_MH_AP_20240416_QE1_6.raw SUBJECT_SAMPLE_FACTORS blank QC3_MDA_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC3_01029_MH_AP_20240416_QE1_12.raw SUBJECT_SAMPLE_FACTORS blank QC4_MDA_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC4_01029_MH_AP_20240416_QE1_18.raw SUBJECT_SAMPLE_FACTORS blank QC5_MDA_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC5_01029_MH_AP_20240416_QE1_25.raw SUBJECT_SAMPLE_FACTORS blank QC6_MDA_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC6_01029_MH_AP_20240416_QE1_26.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_U1_GC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=02_ST_2_G1_11.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_U2_GC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=06_ST_2_G2_8.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_U3_GC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=10_ST_2_G3_13.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_U4_GC Sample source:MDA-MB-231 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=14_ST_2_G4_16.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_S1_GC Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=04_SH_2_g1_15.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_S2_GC Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=08_SH_2_G2_18.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_S3_GC Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=12_SH_2_G3_7.raw SUBJECT_SAMPLE_FACTORS GC-MS MDA_S4_GC Sample source:MDA-MB-231 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=16_SH_2_G4_20.raw SUBJECT_SAMPLE_FACTORS blank QC1_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC1_01028_MH_AP_4.raw SUBJECT_SAMPLE_FACTORS blank QC2_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC2_01028_MH_AP_5.raw SUBJECT_SAMPLE_FACTORS blank QC3_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC3_01028_MH_AP_12.raw SUBJECT_SAMPLE_FACTORS blank QC4_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC4_01028_MH_AP_19.raw SUBJECT_SAMPLE_FACTORS blank QC5_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC5_01028_MH_AP_26.raw SUBJECT_SAMPLE_FACTORS blank QC6_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=QC6_01028_MH_AP_27.raw SUBJECT_SAMPLE_FACTORS blank blank_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=Blank_01028_MH_AP_0.raw SUBJECT_SAMPLE_FACTORS blank Human_Plasma_Reference_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=Human_Plasma_Reference.raw SUBJECT_SAMPLE_FACTORS blank Processing_Blank_MDA_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=Processing_Blank_01028_MH_AP_0.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_U1_LC Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_06.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_U2_LC Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_09.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_U3_LC Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_07.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_U4_LC Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_14.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_S1_LC Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_13.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_S2_LC Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_15.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_S3_LC Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_08.raw SUBJECT_SAMPLE_FACTORS LC-MS PC3_S4_LC Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_12.raw SUBJECT_SAMPLE_FACTORS blank Blank1_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_01.raw SUBJECT_SAMPLE_FACTORS blank Blank2_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_02.raw SUBJECT_SAMPLE_FACTORS blank PB_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_03.raw SUBJECT_SAMPLE_FACTORS blank Blank3_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_04.raw SUBJECT_SAMPLE_FACTORS blank QC1_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_05.raw SUBJECT_SAMPLE_FACTORS blank Blan3_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_10.raw SUBJECT_SAMPLE_FACTORS blank QC2_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_11.raw SUBJECT_SAMPLE_FACTORS blank QC3_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_16.raw SUBJECT_SAMPLE_FACTORS blank Blank4_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_17.raw SUBJECT_SAMPLE_FACTORS blank Blank5_PC3_LC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00327_MH_DM_09132020_LCQE_18.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_U1 Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_05.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_U2 Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_09.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_U3 Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_06.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_U4 Sample source:PC-3 cancer cells | Factor:unsheared RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_08.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_S1 Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_11.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_S2 Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_07.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_S3 Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_12.raw SUBJECT_SAMPLE_FACTORS GC-MS PC3_S4 Sample source:PC-3 cancer cells | Factor:FSS RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_10.raw SUBJECT_SAMPLE_FACTORS blank PC3_Pyridine_blank_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_01.raw SUBJECT_SAMPLE_FACTORS blank Process blank_PC3_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_02.raw SUBJECT_SAMPLE_FACTORS blank QC1_PC3_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_03.raw SUBJECT_SAMPLE_FACTORS blank QC2_PC3_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_04.raw SUBJECT_SAMPLE_FACTORS blank QC3_PC3_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_13.raw SUBJECT_SAMPLE_FACTORS blank QC4_PC3_GC Sample source:blank | Factor:- RAW_FILE_NAME(Raw file name)=00312_MH_DM_20200811_ISQ_14.raw #COLLECTION CO:COLLECTION_SUMMARY PC-3 and MDA-MB-231 cancer cells were grown to 80% in 37C at 5% CO2 and CO:COLLECTION_SUMMARY harvested using 0.05% trypsin. Cells were washed with PBS, resuspended in CO:COLLECTION_SUMMARY serum-free DMEM. Metabolomic profiling was performed on cells that were CO:COLLECTION_SUMMARY processed and snap frozen (using liquid nitrogen) immediately after they were CO:COLLECTION_SUMMARY exposed to 10 pulses of FSS or held in suspension as described in PMID: CO:COLLECTION_SUMMARY 32187555. Specifically, on the final FSS pulse, the cells were transferred into CO:COLLECTION_SUMMARY 10 mL of ice-cold PBS, the number of viable cells was determined, and the cells CO:COLLECTION_SUMMARY were centrifuged at 500g for 3 minutes at 4°C. Cell debris was removed from CO:COLLECTION_SUMMARY pellets by resuspension in ice-cold PBS, 2x106 cells were transferred into a new CO:COLLECTION_SUMMARY tube, the cells were pelleted again, and these washed cell pellets were snap CO:COLLECTION_SUMMARY frozen in liquid nitrogen. CO:SAMPLE_TYPE Cultured cells CO:COLLECTION_METHOD cells were collected in ice cold PBS, centrifuged, then snap frozen using liquid CO:COLLECTION_METHOD nitrogen CO:STORAGE_CONDITIONS -80℃ CO:TISSUE_CELL_QUANTITY_TAKEN 2x10^6 cells #TREATMENT TR:TREATMENT_SUMMARY No additional drug or pharmacological treatments were performed on the PC-3 or TR:TREATMENT_SUMMARY MDA-MB-231 cells prior to FSS exposure. Both cell lines were either held in TR:TREATMENT_SUMMARY suspension or exposure to a syringe and needle pump apparatus for 10 pulses. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolomic profiling was performed on cells that were processed and snap frozen SP:SAMPLEPREP_SUMMARY (using liquid nitrogen) immediately after they were exposed to 10 pulses of FSS SP:SAMPLEPREP_SUMMARY or held in suspension as described above. Specifically, on the final FSS pulse, SP:SAMPLEPREP_SUMMARY the cells were transferred into 10 mL of ice-cold PBS, the number of viable SP:SAMPLEPREP_SUMMARY cells was determined, and the cells were centrifuged at 500g for 3 minutes at SP:SAMPLEPREP_SUMMARY 4°C. Cell debris was removed from pellets by resuspension in ice-cold PBS, SP:SAMPLEPREP_SUMMARY 2x106 cells were transferred into a new tube, the cells were pelleted again, and SP:SAMPLEPREP_SUMMARY these washed cell pellets were snap frozen. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME SeQuant ZIC- pHILIC (150 x 2.1 mm, 5 μm) CH:SOLVENT_A 100% Water; 20 mM ammonium carbonate; 0.1% Ammonium Hydroxide (v/v), pH is ~9.1 CH:SOLVENT_B 100% Acetonitrile CH:FLOW_GRADIENT The gradient starts at 80% B and decreasing to 20% B over 20 minutes; returning CH:FLOW_GRADIENT to 80% B in 0.5 minutes; and held there for 7 minutes. (PMID: 28388410). CH:FLOW_RATE 0.150 mL/min CH:COLUMN_TEMPERATURE 25°C CH:METHODS_FILENAME MetCore_LC-MS_Method_HILIC_082522_Cells.pdf #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS The mass spectrometer was operated in full-scan, polarity-switching mode from 1 MS:MS_COMMENTS to 20 minutes, with the spray voltage set to 3.0 kV, the heated capillary held MS:MS_COMMENTS at 275°C, and the HESI probe held at 350°C. The sheath gas flow was set to 40 MS:MS_COMMENTS units, the auxiliary gas flow was set to 15 units, and the sweep gas flow was MS:MS_COMMENTS set to 1 unit. MS data acquisition was performed in a range of m/z 70–1,000, MS:MS_COMMENTS with the resolution set at 70,000, the AGC target at 1 × 106, and the maximum MS:MS_COMMENTS injection time at 200 ms.(PMID: 28388410) Data Analysis Acquired LC-MS data were MS:MS_COMMENTS processed by Thermo Scientific TraceFinder 4.1 software, and metabolites were MS:MS_COMMENTS identified based on the University of Iowa Metabolomics Core facility MS:MS_COMMENTS standard-confirmed, inhouse library. NOREVA was used for signal drift correction MS:MS_COMMENTS (PMID: 28525573). Data were normalized to the sum of all the measured metabolite MS:MS_COMMENTS ions in that sample. Polarity switching was used as the ion mode. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS pmole/L MS_METABOLITE_DATA_START Samples MDA_U1_LC MDA_U2_LC MDA_U3_LC MDA_U4_LC MDA_S1_LC MDA_S2 MDA_S3_LC MDA_S4_LC PC3_U1_LC PC3_U2_LC PC3_U3_LC PC3_U4_LC PC3_S1_LC PC3_S2_LC PC3_S3_LC PC3_S4_LC Factors Sample source:MDA-MB-231 cancer cells | Factor:unsheared Sample source:MDA-MB-231 cancer cells | Factor:unsheared Sample source:MDA-MB-231 cancer cells | Factor:unsheared Sample source:MDA-MB-231 cancer cells | Factor:unsheared Sample source:MDA-MB-231 cancer cells | Factor:FSS Sample source:MDA-MB-231 cancer cells | Factor:FSS Sample source:MDA-MB-231 cancer cells | Factor:FSS Sample source:MDA-MB-231 cancer cells | Factor:FSS Sample source:PC-3 cancer cells | Factor:unsheared Sample source:PC-3 cancer cells | Factor:unsheared Sample source:PC-3 cancer cells | Factor:unsheared Sample source:PC-3 cancer cells | Factor:unsheared Sample source:PC-3 cancer cells | Factor:FSS Sample source:PC-3 cancer cells | Factor:FSS Sample source:PC-3 cancer cells | Factor:FSS Sample source:PC-3 cancer cells | Factor:FSS 2',3' cyclic AMP 8743.556444 9339.723825 8166.553664 7605.798806 8737.440163 4233.296361 7855.774336 10975.05304 0.565071595 0.267068929 0.828891209 0.40625288 0.715625029 0.684596805 1.707774756 1.71076861 4-Hydroxy-2-nonenal 3023.874036 5251.139076 3648.566448 3375.019083 7371.775322 1912.599361 8963.275695 4576.085661 0 0 0 0 0 0 0 0 3',5' cyclic AMP 7568.269761 12050.92704 14290.295 9096.028316 8766.735309 6258.455502 6189.360162 12891.09065 0.653891427 0.272953045 0.843405015 0.419001716 0.756193551 0.727413385 1.772749351 1.730284987 Acetyl-CoA 4115.477419 4677.929316 6305.035851 3147.786865 15583.99383 2927.012456 4970.600618 6441.648988 0 0 0 0 0 0 0 0 Adenine 4531.761369 1371.880649 2474.67465 864.5969242 9033.143163 2371.708748 9013.860048 2149.069364 0.254911392 0.668641287 0.633278632 0.445877667 1.082011557 0.59028973 0.615317302 0.846958826 Adenosine 9134.910497 7644.510648 9190.131769 3739.44077 8666.212806 3130.672476 7873.683236 5717.567646 0.136713356 1.123074169 0.247122303 0.21460517 2.544645977 0.61447581 0.490337266 0.595519545 Adenosine 3',5'-diphosphate 5291.72706 2510.64533 5776.395076 894.4381678 7830.643829 1866.755208 8315.053503 2301.090479 0.67761327 1.728250277 0.658165574 0.759819019 1.070104626 0.978935391 0.780033936 1.078593112 AMP 3019.899235 2542.093148 3414.162268 3103.668765 3834.874048 1693.337944 3561.059787 3702.543722 0.217831524 1.984315944 0.26675226 0.277621548 2.842535115 1.76380137 0.328091958 0.697986867 Arginine 9695.084961 10860.47376 10469.8864 9265.386414 5417.471793 3795.491735 5299.116964 7762.312869 0 0 0 0 0 0 0 0 ATP 8076.865984 8896.816037 8888.98651 8244.640363 8092.494939 4187.06376 6982.629019 10154.63513 1.269418186 0.813159463 1.330104797 1.015683668 0.304751072 0.419653633 1.138001124 0.810535734 CDP 12749.59623 7991.468091 10764.44829 6697.144415 7191.468569 2386.381567 6534.866563 3106.934378 0.668594052 1.894539638 0.569824791 0.694109491 1.037214518 1.041803915 0.767457337 1.042781949 CMP 7337.987209 8366.7234 9867.008913 10095.24418 5880.183599 3856.016977 5070.898632 8504.145256 0.164600436 2.08146947 0.181535794 0.204381609 2.906478681 1.66016899 0.27887279 0.687809387 CoA 56981.15069 47378.36846 8786.971142 51762.99044 6187.700157 6837.384892 8574.15984 17347.62054 0 0 0 0 0 0 0 0 Creatine 8106.161196 8379.597043 8602.961611 8546.123479 6056.959143 3362.249769 5313.590802 6952.110862 0.80202832 0.812874246 0.960827738 0.808757004 1.146942604 0.831141883 1.1305199 1.180586115 CTP 13527.21222 16297.10782 13833.62769 13417.67193 4475.334982 3531.581093 4182.909018 7133.431748 1.376270371 0.816194406 1.256922096 0.984570697 0.240482153 0.386362263 1.276509604 0.737221985 dAMP 13241.79943 6136.178997 7227.891937 1650.33139 13792.21888 1688.310616 9048.294577 838.9526469 0 0 0 0 0 0 0 0 dATP 17986.46473 20883.36526 25862.25678 19533.96629 3062.901564 8760.692526 4017.212191 3636.286523 1.197822928 0.161269026 1.336849407 0.93950613 0.095575742 0.150849748 1.016546781 1.111360246 dCMP 0.024141459 6116.616303 0.0222504 2900.287017 5442.317881 0.013258932 0.020611565 6118.536179 0.988431 1.202212761 0.804808116 1.047574464 0.496745072 0.264714846 0.889816094 1.003280211 Deoxyadenosine 0 0 0 0 0 0 0 0 0.31270723 0.829076615 0.954019753 0.891485582 0.563484386 0.429201377 1.03654128 0.955650075 dGDP 7928.525815 4573.940651 10745.91494 2252.22335 9699.119084 2973.138972 8834.469107 3566.34686 1.314941935 0.78829361 1.471636208 1.172423678 0.273883901 0.340950442 1.115190853 0.802527179 dGTP 10757.67934 13223.28424 10924.94427 10935.77887 4993.344346 3730.425419 4993.625044 8065.822763 1.295884636 0.853086298 1.391073064 1.063507058 0.296282143 0.408729005 1.112333758 0.817383128 dTDP 0 0 0 0 0 0 0 0 0.43773222 1.764987702 0.239755391 0.416346729 1.206603809 1.078218177 0.544334132 0.797896378 dTTP 8802.133554 13762.02741 10372.4419 10053.49355 3634.806944 2910.453488 4155.890393 6992.900347 1.349037513 0.8181704 1.302374706 1.101116043 0.266406997 0.293981464 0.979061713 0.758073308 FAD 7798.138443 9813.667377 9324.068132 8539.386915 7145.627964 4298.925247 7092.534228 9555.157035 0.89757363 1.048523401 0.933773565 0.998438721 0.699504354 0.538400875 1.018746664 1.074176498 GDP 7621.837073 6795.921829 16583.7598 5648.856074 10125.34208 3804.417782 7270.978784 8953.528429 0.833506786 1.49569206 0.826814121 0.656224579 0.966265144 0.963606303 1.006683037 0.947385854 Glutathione 23353.18828 17511.17307 12280.13168 24597.49486 89.42406016 5134.206393 16252.40408 7430.619544 1.512123148 1.052465448 0.685788798 2.124557299 1.468265047 0.855312357 0.618276186 0.643997744 GMP 9479.979251 3982.923421 9869.418094 2539.144131 11481.03829 4478.225329 10885.29678 5598.160135 0.179190831 1.859006419 0.292006212 0.247291292 3.087487195 1.659861272 0.372622281 0.73823632 GTP 13434.54595 16918.38452 12711.55292 15596.05778 6001.111584 4065.181473 6140.39865 9032.511729 1.522141641 0.78007054 1.31581395 0.730784722 0.265510478 0.452232269 1.481835892 0.671920178 Guanine 2979.464105 4428.193969 3738.788513 1328.877567 12454.3265 7793.47402 14390.55103 10754.02243 0.307782937 1.080761687 0.237460987 0.345926926 2.293791771 1.236008059 0.735736003 1.561470976 Guanosine 6733.801654 2405.248764 1480.865195 1880.110626 14585.44415 10382.66663 6829.880076 7600.462112 0.44243275 0.287403324 0.557327906 0.483939476 0.989910335 0.235823279 1.966613797 2.107331527 Hypoxanthine 5205.137861 5575.04728 6271.228399 2233.835455 9996.666753 6567.907171 14078.92221 9823.654611 0.247687906 0.596007348 0.230747599 0.284438181 2.44993509 1.303534419 0.941969472 1.731313374 IDP 13442.47227 25128.16407 31420.82234 22134.25309 22186.20041 7119.575352 29801.45835 21797.24318 0.641170249 0.76935638 0.524544487 0.901074685 1.079065888 0.448123455 0.696238903 0.930968404 IMP 23224.15224 4411.696651 7415.185832 4146.88424 10963.13673 3129.091692 7495.771056 2404.070184 0.410623996 0.723336924 0.633659965 0.336684155 1.25603039 0.763331811 1.686943502 1.683087524 Inosine 10461.45339 3110.281464 1669.014466 2812.86141 12015.00491 8183.948055 7915.764291 5291.383713 0.354287574 0.129327955 0.532792719 0.343395033 1.438875139 0.291132089 2.253761591 2.161764932 Malondialdehyde 1334.729555 4762.411745 7042.759355 6326.425083 15054.56417 519871.528 6040.939405 3645.013997 0 0 0 0 0 0 0 0 NAD+ 5859.099278 8523.470519 7126.120109 7034.529963 5074.521302 3195.749486 4144.898261 6872.652779 0.897838269 1.020845444 1.015347542 0.976335422 0.713199405 0.693700661 1.037788071 1.055072877 NADH 9146.512209 4977.774119 7179.643686 7094.834112 2202.45608 2580.63789 8599.804419 2509.18837 1.168386044 0.622798828 0.473363322 1.344749296 0.806074004 0.479733181 1.224068065 0.710038791 NADP+ 6559.000559 9354.213951 5620.720033 9379.41404 5082.43089 4309.332231 3973.748038 8531.636439 0.952552736 0.919807096 0.695248689 1.137744478 0.77144269 0.448190762 1.021032021 0.944864551 NADPH 0 0 0 0 0 0 0 0 0.922061575 1.347216151 0.51046658 1.444718293 1.210098911 1.270367438 0.659830795 0.919407455 Oxidized glutathione 1365.762362 5173.998671 6839.833885 913.6603412 7207.168612 2429.709288 1209.746629 6322.552251 0.511985186 1.123534992 1.122579134 0.25004851 0.23229624 0.558963343 1.116724704 1.141313984 PAP 9134.910497 7644.510648 9190.131769 3739.44077 8666.212806 3130.672476 7873.683236 5717.567646 0 0 0 0 0 0 0 0 Thiamine monophosphate 20865.02029 6124.335624 3177.229998 6918.285365 9454.941626 3428.292841 4834.532186 4610.648823 0.554832871 0.908887605 0.499692845 0.583621935 1.138356504 0.649423474 1.39470053 1.529270477 TTP 8802.133554 13762.02741 10372.4419 10053.49355 3634.806944 2910.453488 4155.890393 6992.900347 0 0 0 0 0 0 0 0 Thymine 0 0 0 0 0 0 0 0 0.747007796 0.931104914 0.928801346 0.209771573 0.330255856 0.54242341 1.454589098 0.775618527 UDP 10750.71688 8253.313205 10252.94189 7031.911439 7991.596121 2945.488545 6918.037863 3675.393017 0.66285657 1.783910696 0.598615697 0.769408675 1.056470056 0.903760961 0.700161799 1.03343621 UMP 8491.113645 3132.983714 6679.08917 2619.52741 10238.94123 3612.799299 8757.379399 3278.604189 0.167660036 1.921442735 0.235116514 0.223272593 3.020509608 1.678760763 0.335672258 0.791630127 Uracil 0 0 0 0 0 0 0 0 0.229919465 1.082764624 0.398451659 0.237322259 3.083248685 1.527041672 0.268489101 1.009444866 Uridine 7149.845759 3878.871695 1860.885183 5133.25175 8435.359371 5334.730629 6026.570262 9317.840853 0.584531018 0.831781992 0.457815859 0.606440061 1.451071714 0.629599524 1.33061301 1.64952466 Xanthosine 1482.835457 2564.917188 6907.8251 9324.036313 7468.555383 3818.554288 8514.121921 7240.410518 0.34646746 0.898828849 0.244857703 1.245587225 1.314977306 0.65160365 1.171570126 1.373824086 XMP 4655.724268 4522.141687 8076.057615 6892.574224 14007.81507 2592.390977 14312.68777 2950.673596 0.246069239 0.295355725 0.1334428 0.3522891 1.801056329 0.35911591 1.855640476 1.698750407 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Standardized name Formula Exact mass Super class Main class Sub class Pubchem CID CHEBI_ID HMDB_ID LM_ID KEGG_ID INCHI_KEY REFMET_ID 2',3' cyclic AMP 2',3' cyclic AMP C10H12N5O6P 329.0525 Nucleic acids Purines Purine rNMP 101812 27844 HMDB0011616 C02353 KMYWVDDIPVNLME-KQYNXXCUSA-N RM0033207 4-Hydroxy-2-nonenal - - - - - - - - 3',5' cyclic AMP 3',5' cyclic AMP C10H12N5O6P 329.0525 Nucleic acids Purines Purine rNMP 6076 17489 HMDB0000058 C00575 IVOMOUWHDPKRLL-KQYNXXCUSA-N RM0135874 Acetyl-CoA Acetyl-CoA C23H38N7O17P3S 809.1258 Fatty Acyls Fatty esters Acyl CoAs 444493 15351 HMDB0001206 LMFA07050281 C00024 ZSLZBFCDCINBPY-ZSJPKINUSA-N RM0073686 Adenine Adenine C5H5N5 135.0545 Nucleic acids Purines Aminopurines 190 16708 HMDB0000034 C00147 GFFGJBXGBJISGV-UHFFFAOYSA-N RM0049281 Adenosine Adenosine C10H13N5O4 267.0968 Nucleic acids Purines Purine ribonucleosides 60961 16335 HMDB0000050 C00212 OIRDTQYFTABQOQ-KQYNXXCUSA-N RM0135870 Adenosine 3',5'-diphosphate Adenosine 3',5'-diphosphate C10H15N5O10P2 427.0294 Nucleic acids Purines Purine rNDP 159296 17985 HMDB0000061 C00054 WHTCPDAXWFLDIH-KQYNXXCUSA-N RM0135875 AMP AMP C10H14N5O7P 347.0631 Nucleic acids Purines Purine rNMP 6083 16027 HMDB0000045 C00020 UDMBCSSLTHHNCD-KQYNXXCUSA-N RM0138921 Arginine Arginine C6H14N4O2 174.1117 Organic acids Amino acids and peptides Amino acids 6322 16467 HMDB0000517 C00062 ODKSFYDXXFIFQN-BYPYZUCNSA-N RM0135963 ATP ATP C10H16N5O13P3 506.9958 Nucleic acids Purines Purine rNTP 5957 15422 HMDB0000538 C00002 ZKHQWZAMYRWXGA-KQYNXXCUSA-N RM0138930 CDP CDP C9H15N3O11P2 403.0182 Nucleic acids Pyrimidines Pyrimidine rNDP 6132 17239 HMDB0001546 C00112 ZWIADYZPOWUWEW-XVFCMESISA-N RM0138946 CMP CMP C9H14N3O8P 323.0519 Nucleic acids Pyrimidines Pyrimidine rNMP 6131 17361 HMDB0000095 C00055 IERHLVCPSMICTF-XVFCMESISA-N RM0138923 CoA Coenzyme A C21H36N7O16P3S 767.1152 Nucleic acids Purines Other purines 87642 15346 HMDB0001423 C00010 RGJOEKWQDUBAIZ-IBOSZNHHSA-N RM0157849 Creatine Creatine C4H9N3O2 131.0695 Organic acids Amino acids and peptides Amino acids 586 16919 HMDB0000064 C00300 CVSVTCORWBXHQV-UHFFFAOYSA-N RM0135876 CTP CTP C9H16N3O14P3 482.9845 Nucleic acids Pyrimidines Pyrimidine rNTP 6176 17677 HMDB0000082 C00063 PCDQPRRSZKQHHS-XVFCMESISA-N RM0135881 dAMP dAMP C10H14N5O6P 331.0682 Nucleic acids Purines Purine dNMP 12599 17713 HMDB0000905 C00360 KHWCHTKSEGGWEX-RRKCRQDMSA-N RM0156433 dATP dATP C10H16N5O12P3 491.0008 Nucleic acids Purines Purine dNTP 15993 16284 HMDB0001532 C00131 SUYVUBYJARFZHO-RRKCRQDMSA-N RM0028587 dCMP dCMP C9H14N3O7P 307.0569 Nucleic acids Pyrimidines Pyrimidine dNMP 13945 15918 HMDB0001202 C00239 NCMVOABPESMRCP-SHYZEUOFSA-N RM0136074 Deoxyadenosine Deoxyadenosine C10H13N5O3 251.1018 Nucleic acids Purines Purine deoxyribonucleosides 13730 17256 HMDB0000101 C00559 OLXZPDWKRNYJJZ-RRKCRQDMSA-N RM0049845 dGDP dGDP C10H15N5O10P2 427.0294 Nucleic acids Purines Purine dNDP 135398595 28862 HMDB0000960 C00361 CIKGWCTVFSRMJU-KVQBGUIXSA-N RM0009096 dGTP dGTP C10H16N5O13P3 506.9958 Nucleic acids Purines Purine dNTP 135398599 16497 HMDB0001440 C00286 HAAZLUGHYHWQIW-KVQBGUIXSA-N RM0136101 dTDP dTDP C10H16N2O11P2 402.0229 Nucleic acids Pyrimidines Pyrimidine dNDP 164628 18075 HMDB0001274 C00363 UJLXYODCHAELLY-XLPZGREQSA-N RM0136085 dTTP dTTP C10H17N2O14P3 481.9893 Nucleic acids Pyrimidines Pyrimidine dNTP 64968 18077 HMDB0001342 C00459 NHVNXKFIZYSCEB-XLPZGREQSA-N RM0032432 FAD FAD C27H33N9O15P2 785.1571 Nucleic acids Flavins Flavin nucleotides 643975 16238 HMDB0001248 C00016 VWWQXMAJTJZDQX-UYBVJOGSSA-N RM0138943 GDP GDP C10H15N5O11P2 443.0243 Nucleic acids Purines Purine rNDP 135398619 17552 HMDB0001201 C00035 QGWNDRXFNXRZMB-UUOKFMHZSA-N RM0033168 Glutathione Glutathione C10H17N3O6S 307.0838 Organic acids Amino acids and peptides Tripeptides 124886 16856 HMDB0000125 C00051 RWSXRVCMGQZWBV-WDSKDSINSA-N RM0135889 GMP GMP C10H14N5O8P 363.058 Nucleic acids Purines Purine rNMP 135398631 17345 HMDB0001397 C00144 RQFCJASXJCIDSX-UUOKFMHZSA-N RM0033601 GTP GTP C10H16N5O14P3 522.9907 Nucleic acids Purines Purine rNTP 135398633 15996 HMDB0001273 C00044 XKMLYUALXHKNFT-UUOKFMHZSA-N RM0136084 Guanine Guanine C5H5N5O 151.0494 Nucleic acids Purines Hypoxanthines 135398634 16235 HMDB0000132 C00242 UYTPUPDQBNUYGX-UHFFFAOYSA-N RM0018303 Guanosine Guanosine C10H13N5O5 283.0917 Nucleic acids Purines Purine ribonucleosides 135398635 16750 HMDB0000133 C00387 NYHBQMYGNKIUIF-UUOKFMHZSA-N RM0156423 Hypoxanthine Hypoxanthine C5H4N4O 136.0385 Nucleic acids Purines Hypoxanthines 135398638 17368 HMDB0000157 C00262 FDGQSTZJBFJUBT-UHFFFAOYSA-N RM0135895 IDP IDP C10H14N4O11P2 428.0134 Nucleic acids Purines Purine rNDP 135398651 17808 HMDB0003335 C00104 JPXZQMKKFWMMGK-KQYNXXCUSA-N RM0043289 IMP IMP C10H13N4O8P 348.0471 Nucleic acids Purines Purine rNMP 135398640 17202 HMDB0000175 C00130 GRSZFWQUAKGDAV-KQYNXXCUSA-N RM0000534 Inosine Inosine C10H12N4O5 268.0808 Nucleic acids Purines Purine ribonucleosides 135398641 17596 HMDB0000195 C00294 UGQMRVRMYYASKQ-KQYNXXCUSA-N RM0135908 Malondialdehyde Malondialdehyde C3H4O2 72.0211 Organic oxygen compounds Carbonyl compounds Other carbonyl compounds 10964 566274 HMDB0006112 C19440 WSMYVTOQOOLQHP-UHFFFAOYSA-N RM0156013 NAD+ NAD C21H27N7O14P2 663.1091 Nucleic acids Nicotinamides Nicotinamide dinucleotides 5892 44215 HMDB0000902 C00003 BAWFJGJZGIEFAR-NNYOXOHSSA-N RM0133217 NADH NADH C21H29N7O14P2 665.1248 Nucleic acids Nicotinamides Nicotinamide dinucleotides 439153 16908 HMDB0001487 C00004 BOPGDPNILDQYTO-NNYOXOHSSA-N RM0136399 NADP+ NADP C21H28N7O17P3 743.0755 Nucleic acids Nicotinamides Nicotinamide dinucleotides 5885 44409 HMDB0304435 C00006 XJLXINKUBYWONI-NNYOXOHSSA-N RM0133216 NADPH NADPH C21H30N7O17P3 745.0911 Nucleic acids Nicotinamides Nicotinamide dinucleotides 5884 16474 HMDB0000221 C00005 ACFIXJIJDZMPPO-NNYOXOHSSA-N RM0157850 Oxidized glutathione Oxidized glutathione C20H32N6O12S2 612.152 Organic acids Amino acids and peptides Tripeptides 65359 17858 HMDB0003337 C00127 YPZRWBKMTBYPTK-BJDJZHNGSA-N RM0139022 PAP - - - - - - - - Thiamine monophosphate Thiamine monophosphate C12H17N4O4PS 344.0708 Nucleic acids Pyrimidines Thiamines 3382778 37574 HMDB0002666 C01081 HZSAJDVWZRBGIF-UHFFFAOYSA-N RM0159451 TTP dTTP C10H17N2O14P3 481.9893 Nucleic acids Pyrimidines Pyrimidine dNTP 64968 18077 HMDB0001342 C00459 NHVNXKFIZYSCEB-XLPZGREQSA-N RM0032432 Thymine Thymine C5H6N2O2 126.0429 Nucleic acids Pyrimidines Other pyrimidines 1135 17821 HMDB0000262 C00178 RWQNBRDOKXIBIV-UHFFFAOYSA-N RM0040235 UDP UDP C9H14N2O12P2 404.0022 Nucleic acids Pyrimidines Pyrimidine rNDP 6031 17659 HMDB0000295 C00015 XCCTYIAWTASOJW-XVFCMESISA-N RM0135932 UMP UMP C9H13N2O9P 324.0359 Nucleic acids Pyrimidines Pyrimidine rNMP 6030 16695 HMDB0000288 C00105 DJJCXFVJDGTHFX-XVFCMESISA-N RM0135929 Uracil Uracil C4H4N2O2 112.0273 Nucleic acids Pyrimidines Other pyrimidines 1174 17568 HMDB0000300 C00106 ISAKRJDGNUQOIC-UHFFFAOYSA-N RM0135934 Uridine Uridine C9H12N2O6 244.0695 Nucleic acids Pyrimidines Pyrimidine ribonucleosides 6029 16704 HMDB0000296 C00299 DRTQHJPVMGBUCF-XVFCMESISA-N RM0135933 Xanthosine Xanthosine C10H12N4O6 284.0757 Nucleic acids Purines Purine ribonucleosides 64959 18107 HMDB0000299 UBORTCNDUKBEOP-UUOKFMHZSA-N RM0161017 XMP XMP C10H13N4O9P 364.042 Nucleic acids Purines Purine rNMP 73323 15652 HMDB0001554 C00655 DCTLYFZHFGENCW-UUOKFMHZSA-N RM0010946 METABOLITES_END #END