Inactivation of glycogen synthase kinase-3beta, a downstream target of the raf-1 pathway, is associated with growth suppression in medullary thyroid cancer cells

Mol Cancer Ther. 2007 Mar;6(3):1151-8. doi: 10.1158/1535-7163.MCT-06-0665.

Abstract

Glycogen synthase kinase-3beta (GSK-3beta) is an important regulator of cell proliferation and survival. Conflicting observations have been reported regarding the regulation of GSK-3beta and extracellular signal-regulated kinase (ERK1/2) in cancer cells. In this study, we found that raf-1 activation in human medullary thyroid cancer cells, TT cells, resulted in phosphorylation of GSK-3beta. Inactivation of GSK-3beta in TT cells with well-known GSK-3beta inhibitors such as lithium chloride (LiCl) and SB216763 is associated with both growth suppression and a significant decrease in neuroendocrine markers such as human achaete-scute complex-like 1 and chromogranin A. Growth inhibition by GSK-3beta inactivation was found to be associated with cell cycle arrest due to an increase in the levels of cyclin-dependent kinase inhibitors such as p21, p27, and p15. Additionally, LiCl-treated TT xenograft mice had a significant reduction in tumor volume compared with those treated with control. For the first time, we show that GSK-3beta is a key downstream target of the raf-1 pathway in TT cells. Also, our results show that inactivation of GSK-3beta alone is sufficient to inhibit the growth of TT cells both in vitro and in vivo.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / pharmacology
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Carcinoma, Medullary / enzymology
  • Carcinoma, Medullary / pathology
  • Carcinoma, Medullary / prevention & control*
  • Cell Cycle / drug effects
  • Cell Cycle / physiology
  • Cell Line, Tumor / drug effects
  • Cell Line, Tumor / metabolism
  • Cell Proliferation / drug effects
  • Chromogranin A / metabolism
  • Cyclin D1 / metabolism
  • Cyclin-Dependent Kinase Inhibitor p15 / metabolism
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Cyclin-Dependent Kinase Inhibitor p27 / metabolism
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors*
  • Glycogen Synthase Kinase 3 / metabolism
  • Glycogen Synthase Kinase 3 beta
  • Humans
  • Indoles / pharmacology*
  • Lithium Chloride / pharmacology
  • Maleimides / pharmacology*
  • Mice
  • Mice, Nude
  • NIH 3T3 Cells / drug effects
  • Phosphorylation
  • Proto-Oncogene Proteins c-raf / genetics
  • Proto-Oncogene Proteins c-raf / metabolism*
  • Signal Transduction / drug effects
  • Thyroid Neoplasms / enzymology
  • Thyroid Neoplasms / pathology
  • Thyroid Neoplasms / prevention & control*
  • Xenograft Model Antitumor Assays

Substances

  • Adjuvants, Immunologic
  • Chromogranin A
  • Cyclin-Dependent Kinase Inhibitor p15
  • Cyclin-Dependent Kinase Inhibitor p21
  • Indoles
  • Maleimides
  • SB 216763
  • Cyclin D1
  • Cyclin-Dependent Kinase Inhibitor p27
  • GSK3B protein, human
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, mouse
  • Proto-Oncogene Proteins c-raf
  • Glycogen Synthase Kinase 3
  • Lithium Chloride