UV-induced apoptosis is mediated independent of caspase-9 in MCF-7 cells: a model for cytochrome c resistance

J Biol Chem. 2003 Nov 14;278(46):45793-800. doi: 10.1074/jbc.M307979200. Epub 2003 Sep 3.

Abstract

The importance of the mitochondria in UV-induced apoptosis has become increasingly apparent. Following DNA damage cytochrome c and other pro-apoptotic factors are released from the mitochondria, allowing for formation of the apoptosome and subsequent cleavage and activation of caspase-9. Active caspase-9 then activates downstream caspases-3 and/or -7, which in turn cleave poly(ADP)-ribose polymerase (PARP) and other down-stream targets, resulting in apoptosis. In an effort to understand the mechanisms of Akt-mediated cell survival in breast cancer, we studied the effects of insulin-like growth factor (IGF)-I treatment on UV-treated MCF-7 human breast cancer cells. Apoptosis was induced in MCF-7 cells after UV treatment, as measured by caspase-7 and PARP cleavage, and IGF-I co-treatment protected against this response. Surprisingly caspase-9 cleavage was unchanged with UV and/or IGF-I treatment. Using MCF-7 cells overexpressing caspase-3 we have shown that resistance of caspase-9 to cleavage was not altered by the expression of caspase-3. Furthermore, overexpression of caspase-9 did not enhance PARP or caspase-7 cleavage after UV treatment. Because caspase-8 was activated with UV treatment alone, we believe that UV-induced apoptosis in MCF-7 cells occurs independently of cytochrome c and caspase-9, supporting the existence of a cytoplasmic inhibitor of cytochrome c in MCF-7 cells. We anticipate that such inhibitors may be overexpressed in cancer cells, allowing for treatment resistance.

Publication types

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

MeSH terms

  • Apoptosis*
  • Blotting, Western
  • Breast Neoplasms / metabolism
  • Caspase 3
  • Caspase 7
  • Caspase 9
  • Caspases / metabolism
  • Caspases / physiology*
  • Cell Line
  • Cell Line, Tumor
  • Cell Survival
  • Cytochromes c / metabolism*
  • Enzyme Activation
  • Humans
  • Insulin-Like Growth Factor I / metabolism
  • Microscopy, Fluorescence
  • Phosphatidylinositol 3-Kinases / metabolism
  • Poly(ADP-ribose) Polymerases / metabolism
  • Precipitin Tests
  • Proteins / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA / metabolism
  • Time Factors
  • Transfection
  • Ultraviolet Rays
  • X-Linked Inhibitor of Apoptosis Protein

Substances

  • Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • X-Linked Inhibitor of Apoptosis Protein
  • XIAP protein, human
  • RNA
  • Insulin-Like Growth Factor I
  • Cytochromes c
  • Poly(ADP-ribose) Polymerases
  • Phosphatidylinositol 3-Kinases
  • CASP3 protein, human
  • CASP7 protein, human
  • CASP9 protein, human
  • Caspase 3
  • Caspase 7
  • Caspase 9
  • Caspases