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Phosphatidylcholine metabolism and choline kinase in human osteoblasts
Authors:Zhuo Li  Gengshu Wu  Jelske N van der Veen  Martin HermanssonDennis E Vance
Institution:The Molecular and Cell Biology of Lipids, The Department of Biochemistry, The Alberta Diabetes Institute and Mazankowski Alberta Heart Institute, University of Alberta, Edmonton, Alberta T6G 2S2 Canada
Abstract:There is a paucity of information about phosphatidylcholine (PC) biosynthesis in bone formation. Thus, we characterized PC metabolism in both primary human osteoblasts (HOB) and human osteosarcoma MG-63 cells. Our results show that the CDP-choline pathway is the only de novo route for PC biosynthesis in both HOB and MG-63 cells. Both CK activity and CKα expression in MG-63 cells were significantly higher than those in HOB cells. Silencing of CKα in MG-63 cells had no significant effect on PC concentration but decreased the amount of phosphocholine by approximately 80%. The silencing of CKα also reduced cell proliferation. Moreover, pharmacological inhibition of CK activity impaired the mineralization capacity of MG-63 cells. Our data suggest that CK and its product phosphocholine are required for the normal growth and mineralization of MG-63 cells.
Keywords:ALP  alkaline phosphatase  CK  choline kinase  CT  CTP:phosphocholine cytidylyltransferase  GAPDH  glyceraldehyde-3-phosphate dehydrogenase  HDL  high density lipoproteins  HC-3  hemicholinium-3  HOB  primary human osteoblasts  LDL  low density lipoproteins  LC&ndash  MS  liquid chromatography&ndash  mass spectrometry  PCho  phosphocholine  PC  phosphatidylcholine  PCNA  proliferating cell nuclear antigen  PE  phosphatidylethanolamine  PEMT  phosphatidylethanolamine N-methyltransferase
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