Downregulation of Bcl-2 and activation of caspase-8 in the UVB-induced apoptosis of a cultured human melanoma cell line

Photodermatol Photoimmunol Photomed. 2001 Oct;17(5):218-22. doi: 10.1034/j.1600-0781.2001.170503.x.

Abstract

Purpose: This study was performed to determine the effect of UV radiation on the activation of apoptosis regulatory proteins using cultured human melanoma cells.

Methods: G361 lightly pigmented melanoma cells were irradiated with increasing doses of UVB and analyzed for an apoptotic mechanism using a cell viability test, TEM, FACS, and western blotting analysis.

Results: TEM and FACS showed apoptotic features of cell death after UVB irradiation. Western blotting disclosed downregulation of Bcl-2 and the activation of caspase-9. Caspase-8, a downstream molecule of Fas/FasL interaction, was also activated. The activation of downstream molecules of both caspase-8 and caspase-9 was also demonstrated.

Conclusion: Our data showed that the regulation of the Bcl-2 family and caspase-8 may work together to activate a caspase-3 mediated apoptotic pathway following UVB irradiation of cultured human melanoma cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / radiation effects*
  • Blotting, Western
  • Caspase 8
  • Caspase 9
  • Caspases / metabolism*
  • Chi-Square Distribution
  • Down-Regulation
  • Enzyme Activation / radiation effects
  • Flow Cytometry
  • Humans
  • Melanoma / metabolism
  • Melanoma / pathology*
  • Microscopy, Electron
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • Tumor Cells, Cultured
  • Ultraviolet Rays / adverse effects*

Substances

  • Proto-Oncogene Proteins c-bcl-2
  • CASP8 protein, human
  • CASP9 protein, human
  • Caspase 8
  • Caspase 9
  • Caspases