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Impaired anaplerosis and insulin secretion in insulinoma cells caused by small interfering RNA-mediated suppression of pyruvate carboxylase

dc.contributor.authorNoaman M. Hasanen_US
dc.contributor.authorMelissa J. Longacreen_US
dc.contributor.authorScott W. Stokeren_US
dc.contributor.authorThirajit Boonsaenen_US
dc.contributor.authorSarawut Jitrapakdeeen_US
dc.contributor.authorMindy A. Kendricken_US
dc.contributor.authorJohn C. Wallaceen_US
dc.contributor.authorMichael J. MacDonalden_US
dc.contributor.otherUniversity of Wisconsin School of Medicine and Public Healthen_US
dc.contributor.otherMahidol Universityen_US
dc.contributor.otherUniversity of Adelaideen_US
dc.contributor.otherUniversity of Wisconsin Madisonen_US
dc.date.accessioned2018-07-12T02:16:48Z
dc.date.available2018-07-12T02:16:48Z
dc.date.issued2008-10-17en_US
dc.description.abstractAnaplerosis, the synthesis of citric acid cycle intermediates, by pancreatic beta cell mitochondria has been proposed to be as important for insulin secretion as mitochondrial energy production. However, studies designed to lower the rate of anaplerosis in the beta cell have been inconclusive. To test the hypothesis that anaplerosis is important for insulin secretion, we lowered the activity of pyruvate carboxylase (PC), the major enzyme of anaplerosis in the beta cell. Stable transfection of short hairpin RNA was used to generate a number of INS-1 832/13-derived cell lines with various levels of PC enzyme activity that retained normal levels of control enzymes, insulin content, and glucose oxidation. Glucose-induced insulin release was decreased in proportion to the decrease in PC activity. Insulin release in response to pyruvate alone, 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid (BCH) plus glutamine, or methyl succinate plus β-hydroxybutyrate was also decreased in the PC knockdown cells. Consistent with a block at PC, the most PC-deficient cells showed a metabolic crossover point at PC with increased basal and/or glucose-stimulated pyruvate plus lactate and decreased malate and citrate. In addition, in BCH plus glutamine-stimulated PC knockdown cells, pyruvate plus lactate was increased, whereas citrate was severely decreased, and malate and aspartate were slightly decreased. The incorporation of 14C into lipid from [U-14C]glucose was decreased in the PC knockdown cells. The results confirm the central importance of PC and anaplerosis to generate metabolites from glucose that support insulin secretion and even suggest PC is important for insulin secretion stimulated by noncarbohydrate insulin secretagogues. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc.en_US
dc.identifier.citationJournal of Biological Chemistry. Vol.283, No.42 (2008), 28048-28059en_US
dc.identifier.doi10.1074/jbc.M804170200en_US
dc.identifier.issn1083351Xen_US
dc.identifier.issn00219258en_US
dc.identifier.other2-s2.0-57649118511en_US
dc.identifier.urihttps://repository.li.mahidol.ac.th/handle/20.500.14594/18841
dc.rightsMahidol Universityen_US
dc.rights.holderSCOPUSen_US
dc.source.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=57649118511&origin=inwarden_US
dc.subjectBiochemistry, Genetics and Molecular Biologyen_US
dc.titleImpaired anaplerosis and insulin secretion in insulinoma cells caused by small interfering RNA-mediated suppression of pyruvate carboxylaseen_US
dc.typeArticleen_US
dspace.entity.typePublication
mu.datasource.scopushttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=57649118511&origin=inwarden_US

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