Dataset Information

GNPS - The role of norepinephrine and other metabolites on fallopian tube epithelial metastasis of ovarian cancer

ABSTRACT: DMEM, MOE SCRshRNA, MOE PTENshRNA, MOE PTENshRNAKRAS, MOE PTENshRNAp53shRNA, OVCAR8, OVCAR4 and SKOV3 conditioned media (cell-free media after 4 day incubation of cells with ovary) were collected. These conditioned media samples were lyophilized and metabolites were resuspended with 60:40 ACN:MeOH to obtain 1 mg/mL solution normalized to the dry weight of the extract. An internal standard (rifampicin, 1 uM/mL) was added to samples. Reverse-phase LC was performed on an Elute UHPLC (Bruker Daltonics) using a Phenomonex Kinetex C18 HPLC column (1.7 µm, 2.1 mm x 50 mm, 100 Å) with a sample injection volume of 2 µL. The mobile phase consisted of A (H2O with 0.1% formic acid) and B (ACN with 0.1% formic acid) with a flow rate of 0.5 mL/min. The gradient began with 5% B for 0.3 minutes and was linearly increased to 80% B over 6 minutes. The column was washed using 98% B for 1 minute, and re-equilibrated using 5% B for 1 minute. The temperature of the column oven was 40?. MS spectra were collected using a timsTOF fleX mass spectrometer (Bruker Daltonics) in positive ion mode with a mass range of 50-1000 Da and a spectra rate of 6 Hz. Prior to analysis the instrument was calibrated using 0.5 mM sodium formate. The instrument conditions were set to acquire MS and MS/MS data in separate runs, with 5 MS/MS scans for every 1 MS scan, utilizing a collision energy of 20-50 eV. DMEM, MOE SCRshRNA, MOE PTENshRNA, MOE PTENshRNAKRASG12V, MOE PTENshRNAp53R273H, OVCAR8, OVCAR4 and SKOV3 samples were analyzed in three biological replicates (N=3).

INSTRUMENT(S): timsTOF fleX

ORGANISM(S): Homo Sapiens (ncbitaxon:9606) Mus Musculus (ncbitaxon:10090)

SUBMITTER: Laura Sanchez Joanna Burdette