title: Amino Acid Metabolomics Using LC-MS/MS: Assessment of Cancer-Cell Resistance in a Simulated Tumor Microenvironment.
abstract: We performed a comprehensive quantification of 20 amino acids in RPMI 1640 medium-cultured human colorectal adenocarcinoma cells to evaluate the efficacy of 5-fluorouracil treatment under hypoxic and hypoglycemic conditions, which mimic the tumor microenvironment. In this study, we developed a simple and comprehensive analytical method by using LC-MS/MS connected to the Intrada amino acid column, which eluted amino acids within 9 min. The present method covered a linearity range of 3.6 - 1818 μM, except for Gly (227 - 1818 μM), Ala, Asp, His (7.1 - 1818 μM each), and Trp (3.6 - 909 μM). The limits of detection were in the range of 0.02 - 38.0 pmol per injection in a standard solution. Amino acid concentration data were analyzed using principal-component analysis to represent samples on two-dimensional graphs. Linear discriminant analysis was used to classify samples on the score plots. Using this approach, the effect of 5-fluorouracil treatment could be successfully discriminated at high discrimination rates. Moreover, several amino acids were extracted from corresponding loading plots as candidate markers for distinguishing the effects of the 5-fluorouracil treatment or tumor microenvironmental conditions. These results suggest that our proposed method might be a useful tool for evaluating the efficacy of anticancer drugs in the tumor microenvironment.
authors: Faculty of Pharmaceutical Sciences, Fukuoka University.
authors: Ryoko
authors: R
authors: Tomita
authors: Kenichiro
authors: K
authors: Todoroki
authors: Hiroshi
authors: H
authors: Maruoka
authors: Hideyuki
authors: H
authors: Yoshida
authors: Toshihiro
authors: T
authors: Fujioka
authors: Manabu
authors: M
authors: Nakashima
authors: Masatoshi
authors: M
authors: Yamaguchi
authors: Hitoshi
authors: H
authors: Nohta
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism* / Q000473:pathology; D000596:Amino Acids / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods*; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D019008:Drug Resistance, Neoplasm; D006801:Humans; D057230:Limit of Detection; D055432:Metabolomics; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment
chemical_list: D000596:Amino Acids