title: Development of an Optimized Protocol for NMR Metabolomics Studies of Human Colon Cancer Cell Lines and First Insight from Testing of the Protocol Using DNA G-Quadruplex Ligands as Novel Anti-Cancer Drugs.
abstract: The study of cell lines by nuclear magnetic resonance (NMR) spectroscopy metabolomics represents a powerful tool to understand how the local metabolism and biochemical pathways are influenced by external or internal stimuli. In particular, the use of adherent mammalian cells is emerging in the metabolomics field in order to understand the molecular mechanism of disease progression or, for example, the cellular response to drug treatments. Hereto metabolomics investigations for this kind of cells have generally been limited to mass spectrometry studies. This study proposes an optimized protocol for the analysis of the endo-metabolome of human colon cancer cells (HCT116) by NMR. The protocol includes experimental conditions such as washing, quenching and extraction. In order to test the proposed protocol, it was applied to an exploratory study of cancer cells with and without treatment by anti-cancer drugs, such as DNA G-quadruplex binders and Adriamycin (a traditional anti-cancer drug). The exploratory NMR metabolomics analysis resulted in NMR assignment of all endo-metabolites that could be detected and provided preliminary insights about the biological behavior of the drugs tested.
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", via D. Montesano 49, 80131 Naples, Italy. ilaria.lauri@unina.it.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Lauri
authors.affiliation: Spectroscopy &amp; Chemometrics, Department of Food Science, Faculty of Science, University of Copenhagen, Rolighedsvej 26, 1958 Frederiksberg C, Denmark. frsa@food.ku.dk.|Department of Applied Science and Technology (DISAT), Polytechnic University of Turin-Corso Duca degli Abruzzi 24, 10129 Torino, Italy. frsa@food.ku.dk.
authors.forename: Francesco
authors.initials: F
authors.lastname: Savorani
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", via D. Montesano 49, 80131 Naples, Italy. nunzia.iaccarino@unina.it.
authors.forename: Nunzia
authors.initials: N
authors.lastname: Iaccarino
authors.affiliation: Experimental Chemotherapy Laboratory, Regina Elena National Cancer Institute, 00158 Rome, Italy. zizza@ifo.it.
authors.forename: Pasquale
authors.initials: P
authors.lastname: Zizza
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples "Federico II", via S. Pansini 5, 80131 Naples, Italy. luigimichele.pavone@unina.it.
authors.forename: Luigi Michele
authors.initials: LM
authors.lastname: Pavone
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", via D. Montesano 49, 80131 Naples, Italy. ettore.novellino@unina.it.
authors.forename: Ettore
authors.initials: E
authors.lastname: Novellino
authors.affiliation: Spectroscopy &amp; Chemometrics, Department of Food Science, Faculty of Science, University of Copenhagen, Rolighedsvej 26, 1958 Frederiksberg C, Denmark. se@food.ku.dk.
authors.forename: Søren Balling
authors.initials: SB
authors.lastname: Engelsen
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", via D. Montesano 49, 80131 Naples, Italy. antonio.randazzo@unina.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: Randazzo
keywords: G-quadruplex ligands; Multivariate statistical analysis; NMR spectroscopy; cell metabolomics; colon cancer