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Metabolites 2016, 6(1), 4; doi:10.3390/metabo6010004

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

1
Department of Pharmacy, University of Naples “Federico II”, via D. Montesano 49, 80131 Naples, Italy
2
Spectroscopy & Chemometrics, Department of Food Science, Faculty of Science, University of Copenhagen, Rolighedsvej 26, 1958 Frederiksberg C, Denmark
3
Department of Applied Science and Technology (DISAT), Polytechnic University of Turin—Corso Duca degli Abruzzi 24, 10129 Torino, Italy
4
Experimental Chemotherapy Laboratory, Regina Elena National Cancer Institute, 00158 Rome, Italy
5
Department of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”, via S. Pansini 5, 80131 Naples, Italy
*
Authors to whom correspondence should be addressed.
Academic Editor: Peter Meikle
Received: 25 November 2015 / Revised: 7 January 2016 / Accepted: 11 January 2016 / Published: 15 January 2016
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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. View Full-Text
Keywords: cell metabolomics; colon cancer; NMR spectroscopy; Multivariate statistical analysis; G-quadruplex ligands cell metabolomics; colon cancer; NMR spectroscopy; Multivariate statistical analysis; G-quadruplex ligands
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Lauri, I.; Savorani, F.; Iaccarino, N.; Zizza, P.; Pavone, L.M.; Novellino, E.; Engelsen, S.B.; Randazzo, A. 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. Metabolites 2016, 6, 4.

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