A molecular study of pathways involved in the inhibition of cell proliferation in neuroblastoma B65 cells by the GSK-3 inhibitors lithium and SB-415286

J Cell Mol Med. 2009 Sep;13(9B):3906-17. doi: 10.1111/j.1582-4934.2008.00389.x. Epub 2008 Jun 20.


Pharmacological GSK-3 inhibitors are potential drugs for the treatment of neurodegenerative diseases, cancer and diabetes. We examined the antiproliferative effects of two GSK-3 inhibitors, lithium and SB-415286, on B65 neuroblastoma cell line. Treatment of B65 cells with either drug administered separately caused a decrease in cell proliferation that was associated with G(2)/M cell cycle arrest. Cell-cycle proteins such as cyclins D, E, A, cdk4 and cdk2 were up-regulated. Since lithium and SB-415286-induced G(2)/M arrest we studied changes in the expression of proteins involved in this phase, specifically cyclin B, cdc2 and the phosphorylated form of this protein (tyr15-cdc2). Both drugs increased the expression of tyr15-cdc2, thus inhibiting mitosis. On the other hand, SB-415286 increased the expression of SIRT2, involved in the regulation of proliferation. Moreover, cell-cycle arrest mediated by SB-415286 was accompanied by apoptosis that was not prevented by 100 microM of zVAD-fmk (benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone), a pan-caspase inhibitor. Likewise, GSK-3 inhibitors did not affect the mitochondrial release of apoptosis inducing factor (AIF). We conclude that inhibitors of GSK-3 induced cell-cycle arrest, mediated by the phosphorylation of cdc2 and, in the case of SB-415286, SIRT2 expression, which induced apoptosis in a caspase-independent manner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminophenols / pharmacology*
  • Animals
  • Apoptosis
  • Apoptosis Inducing Factor / metabolism
  • CDC2 Protein Kinase
  • Cell Cycle / drug effects
  • Cell Cycle Proteins / biosynthesis*
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Cell Proliferation
  • Cyclin-Dependent Kinases
  • Enzyme Inhibitors / pharmacology
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors*
  • Lithium / pharmacology*
  • Maleimides / pharmacology*
  • Neuroblastoma / metabolism*
  • Rats
  • Sirtuin 2 / biosynthesis*


  • 3-(3-chloro-4-hydroxyphenylamino)-4-(4-nitrophenyl)-1H-pyrrole-2,5-dione
  • Aminophenols
  • Apoptosis Inducing Factor
  • Cell Cycle Proteins
  • Enzyme Inhibitors
  • Maleimides
  • Sirt2 protein, rat
  • Lithium
  • CDC2 Protein Kinase
  • Cdk1 protein, rat
  • Cyclin-Dependent Kinases
  • Glycogen Synthase Kinase 3
  • Sirtuin 2