Complex regulation of the cyclin-dependent kinase inhibitor p27kip1 in thyroid cancer cells by the PI3K/AKT pathway: regulation of p27kip1 expression and localization

Am J Pathol. 2005 Mar;166(3):737-49. doi: 10.1016/S0002-9440(10)62295-X.

Abstract

Functional inactivation of the tumor suppressor p27(kip1) in human cancer occurs either through loss of expression or through phosphorylation-dependent cytoplasmic sequestration. Here we demonstrate that dysregulation of the PI3K/AKT pathway is important in thyroid carcinogenesis and that p27(kip1) is a key target of the growth-regulatory activity exerted by this pathway in thyroid cancer cells. Using specific PI3K inhibitors (LY294002, wortmannin, and PTEN) and a dominant active AKT construct (myrAKT), we demonstrated that the PI3K/AKT pathway controlled thyroid cell proliferation by regulating the expression and subcellular localization of p27. Results obtained with phospho-specific antibodies and with transfection of nonphosphorylable p27(kip1) mutant constructs demonstrated that PI3K/AKT-dependent regulation of p27(kip1) mislocalization in thyroid cancer cells occurred via phosphorylation of p27(kip1) at T157 and T198 (but not at S10 or T187). Finally, we evaluated whether these results were applicable to human tumors. Analysis of 100 thyroid carcinomas indicated that p27(kip1) phosphorylation at T157/T198 and cytoplasmic mislocalization were preferentially associated with activation of the PI3K/AKT pathway. Thus the PI3/AKT pathway and its effector p27(kip1) play major roles in thyroid carcinogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androstadienes / pharmacology
  • Blotting, Western
  • Bromodeoxyuridine / pharmacology
  • Cell Cycle Proteins / biosynthesis*
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Chromones / pharmacology
  • Cyclin-Dependent Kinase Inhibitor p27
  • Cytoplasm / metabolism
  • Cytosol / metabolism
  • Enzyme Activation
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Microscopy, Fluorescence
  • Morpholines / pharmacology
  • PTEN Phosphohydrolase
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphoric Monoester Hydrolases / metabolism
  • Phosphorylation
  • Protein-Serine-Threonine Kinases / metabolism
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-akt
  • Reverse Transcriptase Polymerase Chain Reaction
  • S-Phase Kinase-Associated Proteins / metabolism
  • Thyroid Neoplasms / metabolism*
  • Time Factors
  • Transfection
  • Tumor Suppressor Proteins / biosynthesis*
  • Tumor Suppressor Proteins / metabolism
  • Wortmannin

Substances

  • Androstadienes
  • Cell Cycle Proteins
  • Chromones
  • Morpholines
  • Proto-Oncogene Proteins
  • S-Phase Kinase-Associated Proteins
  • Tumor Suppressor Proteins
  • Cyclin-Dependent Kinase Inhibitor p27
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • Phosphatidylinositol 3-Kinases
  • AKT1 protein, human
  • Protein-Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-akt
  • Phosphoric Monoester Hydrolases
  • PTEN Phosphohydrolase
  • PTEN protein, human
  • Bromodeoxyuridine
  • Wortmannin