title: Effects of antibiotic antitumor drugs on nucleotide levels in cultured tumor cells: an exploratory method to distinguish the mechanisms of antitumor drug action based on targeted metabolomics.
abstract: Nucleotide pools in mammalian cells change due to the influence of antitumor drugs, which may help in evaluating the drug effect and understanding the mechanism of drug action. In this study, an ion-pair RP-HPLC method was used for a simple, sensitive and simultaneous determination of the levels of 12 nucleotides in mammalian cells treated with antibiotic antitumor drugs (daunorubicin, epirubicin and dactinomycin D). Through the use of this targeted metabolomics approach to find potential biomarkers, UTP and ATP were verified to be the most appropriate biomarkers. Moreover, a holistic statistical approach was put forward to develop a model which could distinguish 4 categories of drugs with different mechanisms of action. This model can be further validated by evaluating drugs with different mechanisms of action. This targeted metabolomics study may provide a novel approach to predict the mechanism of action of antitumor drugs.
authors.affiliation: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Liu
authors.affiliation: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Cuichai
authors.initials: C
authors.lastname: Liu
authors.affiliation: School of Life Science and Bio-pharmaceutics Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Liu
authors.affiliation: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Lixin
authors.initials: L
authors.lastname: Sun
keywords: ADP, adenosine diphosphate; AMP, adenosine monophosphate; ANOVA, analysis of variance; ATP, adenosine triphosphate; AUC, area under the curve; Antibiotic anticancer drugs; CDP, cytidine diphosphate; CTP, cytidine triphosphate; DMEM, Dulbecco׳s modified eagle׳s cell culture media; DMSO, dimethyl sulfoxide; DNA, deoxyribonucleic acid; EC, energy charge; EDTA, ethylene diamine tetra-acetic acid; FCS, fetal calf serum; GDP, guanosine diphosphate; GMP, guanosine monophosphate; GTP, guanosine triphosphate; HEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; Ion-pair HPLC; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Mechanisms of antitumor drug action; Nucleotides; PBS, phosphate buffered saline; PCA, principal component analysis; Potential biomarkers; Principal component analysis; RNA, ribonucleic acid; ROC, receiver operating characteristic; RPMI-1640, Roswell Park Memorial Institute-1640; TBAHS, tetrabutylammonium hydrogen sulfate; TCA, trichloroacetic acid; Targeted metabolomics analysis; Tumor cells; UDP, uridine diphosphate; UTP, uridine triphosphate; dATP, deoxyadenosine triphosphate; dCDP, deoxycytidine diphosphate; dCTP, deoxycytidine triphosphate; dGMP, deoxyribonucleic monophosphate; dGTP, deoxyguanosine triphosphate; dUDP, deoxyuridine diphpsphate; dUTP, deoxyuridine triphosphate