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2016 | 63 | 2 | 247-251
Article title

Glucosylceramide synthase inhibitors D-PDMP and D-EtDO-P4 decrease the GM3 ganglioside level, differ in their effects on insulin receptor autophosphorylation but increase Akt1 kinase phosphorylation in human hepatoma HepG2 cells

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Abstracts
EN
Gangliosides function as modulators of several cell growth related receptors. It was shown for caveolin-rich adipocytes, that GM3 ganglioside binds to insulin receptor (IR), dissociates its complex with caveolin, and thus lowers IR autophosphorylation following insulin treatment. We extended those studies into human hepatocyte-derived HepG2 cells, characterized by a high level of IR but low of caveolin. To lower the glycosphingolipid content, estimated by GM3 concentration, two glucosylceramide synthase inhibitors d-threo-1-pheny-2-decanoylamino-3-morpholino-1-propanol (d-PDMP) and d-threo-1-(3,4,-ethylenedioxy)phenyl-2-palmitoylamino-3-pyrrolidino-1-propanol (d-EtDO-P4) were used. d-PDMP at 40 µM or d-EtDO-P4 at 1 µM concentrations in culture medium decreased the GM3 content to 22.3% (17.8-26.1%) and 18.1% (13.7-24.4%), respectively, of the control value. The reduction of GM3 obtained with d-PDMP was accompanied by a 185.1% (153.5-423.8%) significant increase in the level of IR autophosphorylation following cell stimulation with 100 nM insulin. The effect of d-EtDO-P4 on IR autophosphorylation was smaller amounting to an increase by 134.8% (111.3-167.8%) of the control level and statistically non-significant. The effects of d-PDMP and d-EtDO-P4 could also be detected at the level of Akt1 kinase. In cells grown in the presence of d-PDMP the level of phosphorylated Akt1 was 286.0% (151.4%-621.1%) of that in the control. In this case the effect of d-EtDO-P4 was similar: 223.0% (181.4-315.4%) significant increase in phosphorylated Akt1. We assume that glycosphingolipid depletion in HepG2 cells may affect not only IR autophosphorylation but also, independently, the phosphorylation of Akt1, by modifying the membrane microenvironment of this kinase.
Publisher

Year
Volume
63
Issue
2
Pages
247-251
Physical description
Dates
published
2016
received
2014-11-04
revised
2015-11-18
accepted
2016-03-01
(unknown)
2016-05-12
Contributors
  • Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland
  • Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland
author
  • Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland
  • Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland
References
  • Bieberich E, Freischutz B, Suzuki M, Yu RK (1999) Differential effects of glycolipid biosynthesis inhibitors on ceramide-induced cell death in neuroblastoma cells. J Neurochem 72: 1040-1049. doi: 10.1046/j.1471-4159.1999.0721040.x.
  • Butters TD, Dwek RA, Platt FM (2005) Imino sugar inhibitors for treating lysosomal glycosphingolipidoses. Glycobiology 15: 43R-52R. doi: 10.1093/glycob/cwi076.
  • Chavez JA, Knotts TA, Wang LP, Li G, Dobrowsky RT, Florant GL, Summers SA (2003) A role for ceramide, but not diacylglycerol, in the antagonism of insulin signal transduction by saturated fatty acids. J Biol Chem 278: 10297-10303. doi: 10.1074/jbc.M212307200.
  • Chavez JA, Siddique MM, Wang ST, Ching J, Shayman JA, Summers SA (2014) Ceramides and glucosylceramides are independent antagonists of insulin signaling. J Biol Chem 289: 723-734. doi: 10.1074/jbc.M113.522847.
  • Cinar B, Mukhopadhyay NK, Meng G, Freeman MR (2007) Phosphoinositide 3-kinase-independent non-genomic signals transit from the androgen receptor to Akt1 in membrane rafts microdomains. J Biol Chem 282: 29584-29593. doi: 10.1074/jbc.M703310200.
  • Cohen P (2008) The twentieth century struggle to decipher insulin signaling. Nat Rev Mol Cell Biol 7: 867-873. doi: 10.1038/nrm2043.
  • Cokakli M, Erdal E, Nart D, Yilmaz F, Sagol O, Kilic M, Karademir S, Atabey N (2009) Differential expression of caveolin-1 in hepatocellular carcinoma: correlation with differentiation state, motility and invasion. BMC Cancer 9: 65. doi: 10.1186/1471-2407-9-65
  • Delos Santos RC, Garay C, Antonescu CN (2015) Charming neighborhoods on the cell surface: plasma membrane microdomains regulate receptor tyrosine kinase signaling. Cell Signal 27: 1963-1976. doi: 10.1016/j.cellsig.2015.07.004.
  • di Bartolomeo S, Spinedi A (2001) Differential chemosensitizing effect of two glucosylceramide synthase inhibitors in hepatoma cells. Biochem Biophys Res Commun 288: 269-274. doi: 10.1006/bbrc.2001.5748.
  • Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226: 497-509.
  • Guo S (2014) Insulin signaling, resistance, and metabolic syndrome: insights from mouse models into disease mechanisms. J Endocrinol 220: T1-T23. doi: 10.1530/JOE-13-0327.
  • Inokuchi J, Radin NS (1987) Preparation of the active isomer of 1-phenyl-2-decanoylamino-3-morpholino-1-propanol, inhibitor of murine glucocerebroside synthetase. J Lipid Res 28: 565-571.
  • Inokuchi J (2014) GM3 and diabetes. Glycoconj J 31: 193-197. doi: 10.1007/s10719-013-9516-4.
  • Langeveld M, Aerts JM (2009) Glycosphingolipids and insulin resistance. Prog Lipid Res 48: 196-205. doi: 10.1016/j.plipres.2009.03.002.
  • Lee L, Abe A, Shayman JA (1999) Improved inhibitors of glucosylceramide synthase. J Biol Chem 274: 14662-14669. doi: 10.1074/jbc.274.21.14662.
  • Lopez PH, Schnaar RL (2009) Gangliosides in cell recognition and membrane protein regulation. Curr Opin Struct Biol 19: 549-557. doi: 10.1016/j.sbi.2009.06.001.
  • Neville DC, Coquard V, Priestman DA, te Vruchte DJ, Sillence DJ, Dwek RA, Platt FM, Butters TD (2004) Analysis of fluorescently labeled glycosphingolipid-derived oligosaccharides following ceramide glycanase digestion and anthranilic acid labeling. Analyt Biochem 331: 275-282. doi: 10.1016/j.ab.2004.03.051.
  • Nojiri H, Stroud M, Hakomori S (1991) A specific type of ganglioside as a modulator of insulin-dependent cell growth and insulin receptor tyrosine kinase activity. J Biol Chem 266: 4531-4537.
  • Pacuszka T, Duffard RO, Nishimura RN, Brady RO, Fishman PH (1978) Biosynthesis of bovine thyroid gangliosides. J Biol Chem 253: 5839-5846.
  • Panasiewicz M, Domek H, Hoser G, Kawalec M, Pacuszka T (2003) Structure of the ceramide moiety of GM1 ganglioside determines its occurrence in different detergent-resistant membrane domains in HL-60 cells. Biochemistry 42: 6608-6618. doi: 10.1021/bi0206309.
  • Sandhoff K, Kolter T (2003) Biosynthesis and degradation of mammalian glycosphingolipids. Philos Trans R Soc Lond B Biol Sci 358: 847-861. doi: 10.1098/rstb.2003.1265.
  • Santi SA, Lee H (2010) The Akt isoforms are present at distinct subcellular locations. Am J Physiol Cell Physiol 298: C580-C591. doi: 10.1152/ajpcell.00375.2009.
  • Sasaki A, Hata K, Suzuki S, Sawada M, Wada T, Yamaguchi K, Obinata M, Tateno H, Suzuki H, Miyagi T (2003) Overexpression of plasma membrane-associated sialidase attenuates insulin signaling in transgenic mice. J Biol Chem 278: 27896-27902. doi: 10.1074/jbc.M212200200.
  • Schnaar RL (1994) Isolation of glycosphingolipids. Methods Enzymol 230: 348-370.
  • Schultze SM, Jensen J, Hemmings BA, Tschopp O, Niessen M (2011) Promiscuous affairs of PKB/AKT in metabolism. Arch Physiol Biochem 117: 70-77. doi: 10.3109/13813455.2010.539236.
  • Schwarzmann G, Sandhoff K (1987) Lysogangliosides: synthesis and use in preparing labeled gangliosides. Methods Enzymol 138: 319-341.
  • Spitalnik PF, Danley JM, Burger SR, Spitalnik SL (1989) The glycosphingolipid composition of the human hepatoma cell line, HepG2. Arch Biochem Biophys 273: 578-591.
  • Stralfors P (2012) Caveolins and caveolae, roles in insulin signalling and diabetes. Adv Exp Med Biol 729: 111-126. doi: 10.1007/978-1-4614-1222-9_8.
  • Svennerholm L, Fredman P (1980) A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617: 97-109.
  • Vainio S, Heino S, Månsson JE, Fredman P, Kuismanen E, Vaarala O, Ikonen E (2002) Dynamic association of human insulin receptor with lipid rafts in cells lacking caveolae. EMBO Rep 3: 95-100. doi: 10.1093/embo-reports/kvf010.
  • Yamashita Y, Hashiramoto A, Haluzik M, Mizukami H, Beck S, Norton A, Kono M, Tsuji S, Daniotti JL, Werth N, Sandhoff R, Sandhoff K, Proia RL (2003) Enhanced insulin sensitivity in mice lacking ganglioside GM3. Proc Nat Acad Sci USA 100: 3445-3498. doi: 10.1073/pnas.0635898100.
Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv63p247kz
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