Constitutive cellular expression of PI 3-kinase is distinct from transient expression

Biochem Biophys Res Commun. 2000 Jun 16;272(3):822-9. doi: 10.1006/bbrc.2000.2806.

Abstract

The discovery that the PTEN tumor suppressor encodes a phosphoinositide 3-phosphatase has raised interest in the effects of constitutive activation of PI 3-kinase. To gain insight into PI 3-kinase function, we have stably expressed a myristoylated form of the catalytic subunit p110alpha (myr-p110) in cells. The myr-p110 associated with the endogenous p85 regulatory subunit and retained lipid and protein kinase activity. Stable lines expressing myr-p110 had 2- to 4-fold more PI 3-kinase activity than controls. Expression of myr-p110 altered cellular morphology and increased the saturation density in culture. These clones were morphologically transformed but Akt and pp70(s6k) were not constitutively activated in contrast to transient assays and from tumor cell lines deficient in PTEN. In addition, the ability of PDGF to induce activation of Akt and pp70(s6k) was diminished. Therefore, expression of a myristoylated PI 3-kinase in murine fibroblasts induces a morphological transformation of the cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Actins / metabolism
  • Animals
  • Catalytic Domain / genetics
  • Catalytic Domain / physiology
  • Cell Adhesion
  • Cell Count
  • Cell Line, Transformed
  • Cell Size
  • Cell Transformation, Neoplastic*
  • Contact Inhibition
  • Enzyme Activation / drug effects
  • Gene Expression*
  • Mice
  • Molecular Weight
  • Myristic Acid / metabolism
  • PTEN Phosphohydrolase
  • Phosphatidylinositol 3-Kinases / chemistry
  • Phosphatidylinositol 3-Kinases / genetics*
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphoric Monoester Hydrolases / deficiency
  • Phosphoric Monoester Hydrolases / genetics
  • Phosphoric Monoester Hydrolases / metabolism
  • Platelet-Derived Growth Factor / pharmacology
  • Precipitin Tests
  • Protein-Serine-Threonine Kinases*
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-akt
  • Ribosomal Protein S6 Kinases / metabolism
  • Signal Transduction / drug effects
  • Transfection
  • Tumor Suppressor Proteins*

Substances

  • Actins
  • Platelet-Derived Growth Factor
  • Proto-Oncogene Proteins
  • Tumor Suppressor Proteins
  • Myristic Acid
  • Phosphatidylinositol 3-Kinases
  • Protein-Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-akt
  • Ribosomal Protein S6 Kinases
  • Phosphoric Monoester Hydrolases
  • PTEN Phosphohydrolase