title: Alterations of the exo- and endometabolite profiles in breast cancer cell lines: A mass spectrometry-based metabolomics approach.
abstract: In recent years, knowledge about metabolite changes which are characteristic for the physiologic state of cancer cells has been acquired by liquid chromatography coupled to mass spectrometry. Distinct molecularly characterized breast cancer cell lines provide an unbiased and standardized in vitro tumor model reflecting the heterogeneity of the disease. Tandem mass spectrometry is a widely applied analytical platform and highly sensitive technique for analysis of complex biological samples. Endo- and exometabolite analysis of the breast cancer cell lines MDA-MB-231, -453 and BT-474 as well as the breast epithelial cell line MCF-10A has been performed using two different analytical platforms: UPLC-ESI-Q-TOF based on a scheduled precursor list has been applied for highlighting of significant differences between cell lines and HPLC-ESI-QqQ using multiple reaction monitoring has been utilized for a targeted approach focusing on RNA metabolism and interconnected pathways, respectively. Statistical analysis enabled a clear discrimination of the breast epithelial from the breast cancer cell lines. As an effect of oxidative stress, a decreased GSH/GSSG ratio has been detected in breast cancer cell lines. The triple negative breast cancer cell line MDA-MB-231 showed an elevation in nicotinamide, 1-ribosyl-nicotinamide and NAD+ reflecting the increased energy demand in triple negative breast cancer, which has a more aggressive clinical course than other forms of breast cancer. Obtained distinct metabolite pattern could be correlated with distinct molecular characteristics of breast cancer cells. Results and methodology of this preliminary in vitro study could be transferred to in vivo studies with breast cancer patients.
authors.affiliation: Center for Biological Systems Analysis ZBSA, Albert-Ludwigs-University Freiburg, 79104 Freiburg, Germany; Institute of Biology II, Albert-Ludwigs-University Freiburg, 79104 Freiburg, Germany. Electronic address: lucas.willmann@mail.zbsa.uni-freiburg.de.
authors.forename: Lucas
authors.initials: L
authors.lastname: Willmann
authors.affiliation: Center for Biological Systems Analysis ZBSA, Albert-Ludwigs-University Freiburg, 79104 Freiburg, Germany; Institute of Biology II, Albert-Ludwigs-University Freiburg, 79104 Freiburg, Germany; Spemann Graduate School of Biology and Medicine, Albert-Ludwigs-University Freiburg, Germany. Electronic address: manuel.schlimpert@sgbm.uni-freiburg.de.
authors.forename: Manuel
authors.initials: M
authors.lastname: Schlimpert
authors.affiliation: Department of Obstetrics and Gynecology, University Hospital Freiburg, Freiburg 79106, Germany; German Cancer Consortium (DKTK), Heidelberg, Germany; German Cancer Research Center (DKFZ), Heidelberg, Germany. Electronic address: marc.hirschfeld@uniklinik-freiburg.de.
authors.forename: Marc
authors.initials: M
authors.lastname: Hirschfeld
authors.affiliation: University Medical Center Freiburg, 79106 Freiburg, Germany. Electronic address: thalia.erbes@uniklinik-freiburg.de.
authors.forename: Thalia
authors.initials: T
authors.lastname: Erbes
authors.affiliation: Universitätsfrauenklinik der Heinrich-Heine-Universität Düsseldorf, 40225 Düsseldorf, Germany. Electronic address: Hans.Neubauer@med.uni-duesseldorf.de.
authors.forename: Hans
authors.initials: H
authors.lastname: Neubauer
authors.affiliation: Department of Obstetrics and Gynecology, RWTH Aachen University, 52074 Aachen, Germany. Electronic address: estickeler@ukaachen.de.
authors.forename: Elmar
authors.initials: E
authors.lastname: Stickeler
authors.affiliation: Center for Biological Systems Analysis ZBSA, Albert-Ludwigs-University Freiburg, 79104 Freiburg, Germany. Electronic address: Bernd.Kammerer@zbsa.uni-freiburg.de.
authors.forename: Bernd
authors.initials: B
authors.lastname: Kammerer
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D016000:Cluster Analysis; D005260:Female; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D055432:Metabolomics
keywords: Breast cancer; Liquid chromatography; Mass spectrometry; Metabolomics; Quadrupole time-of-flight