title: Analysis of deoxyribonucleotide pools in human cancer cell lines using a liquid chromatography coupled with tandem mass spectrometry technique.
abstract: Endogenous ribonucleotides and deoxyribonucleotides play a critical role in cell function, and determination of their levels is of fundamental importance in understanding key cellular processes involved in energy metabolism and molecular and biochemical signaling pathways. In this study, we determined the respective ribonucleotide and deoxyribonucleotide pool sizes in different human cell lines using a simple sample preparation method and LC/MS/MS. This assay was used to determine alterations in deoxyribonucleotide pools in human pancreatic PANC1 cells in response to hypoxia and to treatment with either hydroxyurea or aphidicolin. The levels of all deoxyribonucleotide metabolites decreased with hypoxia treatment, except for dUMP, which increased by two-fold. This LC/MS/MS assay is simple, fast, and sensitive, and it represents a significant advance over previously published methodologies.
authors.affiliation: Department of Pharmacology, Section of Medical Oncology, Yale School of Medicine, New Haven, CT 06520, USA. wei.zhang.wz87@yale.edu
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Shenglan
authors.initials: S
authors.lastname: Tan
authors.forename: Elijah
authors.initials: E
authors.lastname: Paintsil
authors.forename: Ginger E
authors.initials: GE
authors.lastname: Dutschman
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Gullen
authors.forename: Edward
authors.initials: E
authors.lastname: Chu
authors.forename: Yung-Chi
authors.initials: YC
authors.lastname: Cheng
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000737:chemistry; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D003854:Deoxyribonucleotides / Q000737:chemistry* / Q000302:isolation & purification; D056945:Hep G2 Cells; D006801:Humans; D015458:Leukemia, T-Cell / Q000378:metabolism; D008113:Liver Neoplasms / Q000737:chemistry; D008175:Lung Neoplasms / Q000737:chemistry; D010190:Pancreatic Neoplasms / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards
chemical_list: D003854:Deoxyribonucleotides