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

• Authors: Natalia Fedoryszak-Kuśka , Mirosława Panasiewicz , Hanna Domek , Tadeusz Pacuszka
• Languages of publication: EN

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.

Keywords

EN

Akt1; GM3 ganglioside; glycosphingolipid depletion; insulin receptor

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2016
• Volume: 63
• Issue: 2
• Pages: 247-251

Dates

published

2016

received

2014-11-04

revised

2015-11-18

accepted

2016-03-01

(unknown)

2016-05-12

Contributors

author

Natalia Fedoryszak-Kuśka

• Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland

author

Mirosława Panasiewicz

• Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland

author

Hanna Domek

• Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland

author

Tadeusz Pacuszka

• Department of Biochemistry and Molecular Biology, Medical Center of Postgraduate Education, Warsaw, Poland

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Identifiers

DOI

10.18388/abp.2014_930