Protein kinase Cdelta overexpression enhances radiation sensitivity via extracellular regulated protein kinase 1/2 activation, abolishing the radiation-induced G(2)-M arrest

Cell Growth Differ. 2002 May;13(5):237-46.

Abstract

Protein kinase C (PKC) has been widely implicated in regulation ofcell growth/cell cycle progression and apoptosis. However,the role of PKCdelta in radiosensitivity and cell cycle regulation remains unclear. Overexpression of PKCdelta increased Ca2+-independent PKC activity without altering other PKC isoforms (PKCalpha, -beta1, -epsilon, and -zeta), and extracellular regulated protein kinase (ERK) 1/2 activity was also increased in PKCdelta-specific manner. A clonogenic survival assay showed that PKCdelta-overexpressed cells had more radiosensitivity and pronounced induction of apoptosis than control cells. Flow cytometric analysis revealed that PKCdelta made the cells escape from radiation-induced G(2)-M arrest. Moreover, p53 and p21(Waf) induction by radiation were higher in PKCdelta-overexpressed cells than control cells, and PKCdelta-mediated apoptosis was reduced, when radiation-induced ERK1/2 activity was inhibited by PD98059. Furthermore, PKCdelta antisense and rottlerin, PKC inhibitor-abrogated PKCdelta-mediated radiosensitivity and reduced ERK1/2 activity to the control vector level. These results demonstrated that PKCdelta overexpression enhanced radiation-induced apoptosis and radiosensitivity via ERK1/2 activation, thereby abolishing the radiation-induced G(2)-M arrest and finally apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Acetophenones / pharmacology
  • Animals
  • Antisense Elements (Genetics)
  • Apoptosis / physiology
  • Apoptosis / radiation effects
  • Benzopyrans / pharmacology
  • Colony-Forming Units Assay
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins / genetics
  • Enzyme Inhibitors / pharmacology
  • G2 Phase / physiology*
  • G2 Phase / radiation effects
  • Gene Expression Regulation, Enzymologic
  • Isoenzymes / antagonists & inhibitors
  • Isoenzymes / genetics*
  • Isoenzymes / metabolism
  • Mice
  • Mitogen-Activated Protein Kinase 1 / metabolism*
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases / metabolism*
  • Mitosis / physiology*
  • Mitosis / radiation effects
  • Phosphoproteins / metabolism
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / genetics*
  • Protein Kinase C / metabolism
  • Protein Kinase C-delta
  • Radiation Tolerance / physiology

Substances

  • Acetophenones
  • Antisense Elements (Genetics)
  • Benzopyrans
  • Cdkn1a protein, mouse
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins
  • Enzyme Inhibitors
  • Isoenzymes
  • Phosphoproteins
  • rottlerin
  • Prkcd protein, mouse
  • Protein Kinase C
  • Protein Kinase C-delta
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases