title: Characterization of oncogene-induced metabolic alterations in hepatic cells by using ultrahigh performance liquid chromatography-tandem mass spectrometry.
abstract: Elucidation of altered metabolic pathways by using metabolomics may open new avenues for basic research on disease mechanisms and facilitate the development of novel therapeutic strategies. Here, we report the development of ultrahigh performance liquid chromatography-tandem mass spectrometry-based metabolomics platform with capability of measuring both cationic and anionic intermediates in cellular metabolism. The platform was established based on the hydrophobic ion-pairing interaction chromatography coupled with tandem mass spectrometry in multiple reaction monitoring (MRM) mode. The MRM transitions were created and optimized via energy-resolved collision-induced dissociation experiments, serving as an essential reference point for the quantification and identification. For chromatographic separation, application of hydrophobic ion-pairing interaction led to dramatic enhancement on retention of water-soluble metabolites and provision of good peak shapes. Two volatile ion-pairing reagents, namely heptafluorobutyric acid and tributylamine, were used with dedicated C18 columns as complementary separation systems coupled with the MRM analysis, allowing measurement of the metabolites of interest at nanomolar levels. The developed platform was successfully applied to investigate the altered metabolism in hepatic cells with over-expression of an oncogene, thus can provide important information on the rewired metabolism.
authors.affiliation: State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Hong Kong SAR, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Tang
authors.affiliation: Department of Clinical Oncology, The University of Hong Kong, Hong Kong SAR, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Cao
authors.affiliation: State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Hong Kong SAR, China.
authors.forename: Shuhai
authors.initials: S
authors.lastname: Lin
authors.affiliation: Shenzhen Key Laboratory of Translational Medicine of Tumor and Cancer Research Centre, School of Medicine, Shenzhen University, Shenzhen, China.
authors.forename: Li
authors.initials: L
authors.lastname: Fu
authors.affiliation: State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Hong Kong SAR, China.
authors.forename: Shangfu
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Clinical Oncology, The University of Hong Kong, Hong Kong SAR, China.
authors.forename: Xin-Yuan
authors.initials: XY
authors.lastname: Guan
authors.affiliation: State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Hong Kong SAR, China. Electronic address: zwcai@hkbu.edu.hk.
authors.forename: Zongwei
authors.initials: Z
authors.lastname: Cai
mesh_terms: D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D055432:Metabolomics / Q000379:methods*; D009857:Oncogenes / Q000235:genetics*; D010448:Peptide Initiation Factors / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000097574:Eukaryotic Translation Initiation Factor 5A
chemical_list: D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D000097574:Eukaryotic Translation Initiation Factor 5A
keywords: Cancer metabolism; Hydrophobic ion-pairing interaction; Tandem mass spectrometry; Targeted metabolomics