Insulin receptor substrate-1 and -2 mediate resistance to glucose-induced caspase-3 activation in human neuroblastoma cells

• Published in: Biochimica et biophysica acta Vol. 1812; no. 5; pp. 573 - 580
• Main Authors: Stöhr, Oliver, Hahn, Johann, Moll, Lorna, Leeser, Uschi, Freude, Susanna, Bernard, Corinna, Schilbach, Katharina, Markl, Andreas, Udelhoven, Michael, Krone, Wilhelm, Schubert, Markus
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-05-2011; Elsevier
• Subjects: Apoptosis; Apoptosis - drug effects; Apoptosis Regulatory Proteins - genetics; Apoptosis Regulatory Proteins - metabolism; Bcl-2-Like Protein 11; bcl-Associated Death Protein - genetics; bcl-Associated Death Protein - metabolism; Biochemistry, Molecular Biology; Blotting, Western; Caspase 3 - metabolism; Enzyme Activation; Forkhead Box Protein O1; Forkhead Transcription Factors - genetics; Forkhead Transcription Factors - metabolism; Glucose; Glucose - pharmacology; Humans; Immunoprecipitation; Insulin receptor substrate; Insulin Receptor Substrate Proteins - genetics; Insulin Receptor Substrate Proteins - metabolism; Life Sciences; Membrane Proteins - genetics; Membrane Proteins - metabolism; MnSOD; Neuroblastoma - drug therapy; Neuroblastoma - metabolism; Neuroblastoma - pathology; Neuron; Phosphatidylinositol 3-Kinases - metabolism; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - metabolism; Proto-Oncogene Proteins c-akt; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; Signal Transduction - drug effects; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism; Sweetening Agents - pharmacology; Tumor Cells, Cultured; Neuron; Glucose; Insulin receptor substrate; MnSOD; Apoptosis; neuron; apoptosis; glucose; insulin receptor substrate
• ISSN: 0925-4439; 0006-3002; 1879-260X
• DOI: 10.1016/j.bbadis.2011.02.006

Hyperglycemia in patients with type 2 diabetes causes multiple neuronal complications, e.g., diabetic polyneuropathy, cognitive decline, and embryonic neural crest defects due to increased apoptosis. Possible mechanisms of neuronal response to increased glucose burden are still a matter of debate. Insulin and insulin-like growth factor-1 (IGF-1) receptor signaling inhibits glucose-induced caspase-3 activation and apoptotic cell death. The insulin receptor substrates (IRS) are intracellular adapter proteins mediating insulin's and IGF-1's intracellular effects. Even though all IRS proteins have similar function and structure, recent data suggest different actions of IRS-1 and IRS-2 in mediating their anti-apoptotic effects in glucose neurotoxicity. We therefore investigated the role of IRS-1/-2 in glucose-induced caspase-3 activation using human neuroblastoma cells. Overexpression of IRS-1 or IRS-2 caused complete resistance to glucose-induced caspase-3 cleavage. Inhibition of PI3-kinase reversed this protective effect of IRS-1 or IRS-2. However, MAP-kinases inhibition had only minor impact. IRS overexpression increased MnSOD abundance as well as BAD phosphorylation while Bim and BAX levels remained unchanged. Since Akt promotes cell survival at least partially via phosphorylation and inhibition of downstream forkhead box-O (FoxO) transcription factors, we generated neuroblastoma cells stably overexpressing a dominant negative mutant of FoxO1 mimicking activation of the insulin/IGF-1 pathway on FoxO-mediated transcription. Using these cells we showed that FoxO1 is not involved in neuronal protection mediated by increased IRS-1/-2 expression. Thus, overexpression of both IRS-1 and IRS-2 induces complete resistance to glucose-induced caspase-3 activation via PI3-kinase mediated BAD phosphorylation and MnSOD expression independent of FoxO1.
► Insulin receptor substrate-1 and -2 mediate resistance to glucose-induced caspase-3 activation in human neuroblastoma cells. ► IRS-1/-2 overexpression induces resistance to glucose-induced caspase-3 activation. ► Inhibition of PI3-kinase reverses the protective effect of IRS-1/-2. ► FoxO1 is not involved in neuronal protection mediated by increased IRS-1/-2 expression. ► IRS-1/-2 regulates neuronal MnSOD expression.