title: Inhibition of transketolase by oxythiamine altered dynamics of protein signals in pancreatic cancer cells.
abstract: Oxythiamine (OT), an analogue of anti-metabolite, can suppress the nonoxidative synthesis of ribose and induce cell apoptosis by causing a G1 phase arrest in vitro and in vivo. However, the molecular mechanism remains unclear yet. In the present study, a quantitative proteomic analysis using the modified SILAC method (mSILAC) was performed to determine the effect of metabolic inhibition on dynamic changes of protein expression in MIA PaCa-2 cancer cells treated with OT at various doses (0 μM, 5 μM, 50 μM and 500 μM) and time points (0 h, 12 h and 48 h). A total of 52 differential proteins in MIA PaCa-2 cells treated with OT were identified, including 14 phosphorylated proteins. Based on the dynamic expression pattern, these proteins were categorized in three clusters, straight down-regulation (cluster 1, 37% of total proteins), upright "V" shape expression pattern (cluster 2, 47.8% total), and downright "V" shape pattern (cluster 3, 15.2% total). Among them, Annexin A1 expression was significantly down-regulated by OT treatment in time-dependent manner, while no change of this protein was observed in OT dose-dependent fashion. Pathway analysis suggested that inhibition of transketolase resulted in changes of multiple cellular signaling pathways associated with cell apoptosis. The temporal expression patterns of proteins revealed that OT altered dynamics of protein expression in time-dependent fashion by suppressing phosphor kinase expression, resulting in cancer cell apoptosis. Results from this study suggest that interference of single metabolic enzyme activity altered multiple cellular signaling pathways.
authors.affiliation: Genomics & Functional Proteomics Laboratories, Osteoporosis Research Center, Creighton University Medical Center, 601 N 30th ST, Suite 6730, Omaha, NE 68131, USA.|Department of Respiratory Medicine, The Fifth Hospital of Dalian, Dalian 116027, China.
authors.forename: Jiarui
authors.initials: J
authors.lastname: Wang
authors.affiliation: The Medical College of Dalian University, Dalian Economic & Technological Development Zone, Dalian 116622, China.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Genomics & Functional Proteomics Laboratories, Osteoporosis Research Center, Creighton University Medical Center, 601 N 30th ST, Suite 6730, Omaha, NE 68131, USA.
authors.forename: Danjun
authors.initials: D
authors.lastname: Ma
authors.affiliation: Metabolomics Core, UCLA Center of Excellence in Pancreatic Diseases, Harbor-UCLA Medical Center, Torrance, CA 90502, USA.
authors.forename: Wai-Nang Paul
authors.initials: WP
authors.lastname: Lee
authors.affiliation: Genomics & Functional Proteomics Laboratories, Osteoporosis Research Center, Creighton University Medical Center, 601 N 30th ST, Suite 6730, Omaha, NE 68131, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Xiao
authors.affiliation: Genomics & Functional Proteomics Laboratories, Osteoporosis Research Center, Creighton University Medical Center, 601 N 30th ST, Suite 6730, Omaha, NE 68131, USA.
authors.forename: Yingchun
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Metabolomics Core, UCLA Center of Excellence in Pancreatic Diseases, Harbor-UCLA Medical Center, Torrance, CA 90502, USA.
authors.forename: Vay Liang
authors.initials: VL
authors.lastname: Go
authors.affiliation: Department of Respiratory Medicine, Dalian Medical University, Dalian 116027, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Molecular Clinical Pharmacology, City of Hope Cancer Center, Duarte, CA 90101, USA.
authors.forename: Yun
authors.initials: Y
authors.lastname: Yen
authors.affiliation: Genomics & Functional Proteomics Laboratories, Osteoporosis Research Center, Creighton University Medical Center, 601 N 30th ST, Suite 6730, Omaha, NE 68131, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Recker
authors.affiliation: Genomics & Functional Proteomics Laboratories, Osteoporosis Research Center, Creighton University Medical Center, 601 N 30th ST, Suite 6730, Omaha, NE 68131, USA.
authors.forename: Gary Guishan
authors.initials: GG
authors.lastname: Xiao
keywords: 15 N stable isotope; Metabolic inhibitor; Metabolic therapy; Oxythiamine; Pancreas cancer; Phosphorylation; Quantitative proteomics; Transketolase; Turnover rate
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