Amino Acid Metabolomics Using LC-MS/MS: Assessment of Cancer-Cell Resistance in a Simulated Tumor Microenvironment

• TOMITA Ryoko

  Faculty of Pharmaceutical Sciences, Fukuoka University

• TODOROKI Kenichiro

  Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka

• MARUOKA Hiroshi

  Faculty of Pharmaceutical Sciences, Fukuoka University

• YOSHIDA Hideyuki

  Faculty of Pharmaceutical Sciences, Fukuoka University

• FUJIOKA Toshihiro

  Faculty of Pharmaceutical Sciences, Fukuoka University

• NAKASHIMA Manabu

  Faculty of Pharmaceutical Sciences, Fukuoka University

• YAMAGUCHI Masatoshi

  Faculty of Pharmaceutical Sciences, Fukuoka University

• NOHTA Hitoshi

  Faculty of Pharmaceutical Sciences, Fukuoka University

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.

Journal

• Analytical Sciences

  Analytical Sciences 32 (8), 893-900, 2016

  The Japan Society for Analytical Chemistry

Keywords

• Amino acid metabolomics
• liquid chromatography–tandem mass spectrometry
• 5-fluorouracil
• hypoxia
• hypoglycemia

Details

• CRID: 1390001204259636224
• NII Article ID: 130005256428
• NII Book ID: AA10500785
• DOI: 10.2116/analsci.32.893
• ISSN: 13482246
• ISSN: 09106340
• NDL BIB ID: 027552984
• PubMed: 27506717
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I027552984
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.32.893.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.32.893/fulltext.html
• Web Site: https://www.jstage.jst.go.jp/article/analsci/32/8/32_893/_pdf
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed