Targeted metabolic analysis of nucleotides and identification of biomarkers associated with cancer in cultured cell models

Abstract

Cancer, like other diseases accompanied by metabolic changes, shows characteristic DNA/RNA modifications and activities of modifying enzymes, resulting in fluctuations in nucleoside levels. In this study, we undertook targeted metabolomic analyses of nucleotides in different cancer cell culture models using a sensitive and reproducible ion-pair HPLC method. The experimental data were analyzed by principal component analysis (PCA) to identify potential biomarkers in cancer cells, and statistical significance was determined by one-way analysis of variance. As a result, a clear differentiation of normal and tumor cells into two clusters was shown, indicating abnormal metabolism of nucleotides in tumor cells. Six variables (AMP, UDP, CTP levels with a significance of PUTP (0.938)>CTP=GMP (0.896)>AMP (0.812)>UDP (0.792), so we conclude that ATP and UTP are the best potential biomarkers in tumor cells. This study may provide a valuable tool for studying minute alterations of intracellular nucleotide pools induced by anticancer/antiviral drugs, diseases or environmental factors.

Keywords

• Ion-pair HPLC
• Tumor cells
• Nucleotides
• Targeted metabolomics analysis
• PCA