Apoptosis induced by ultraviolet B in HPV-immortalized human keratinocytes requires caspase-9 and is death receptor independent

Exp Dermatol. 2006 Jan;15(1):23-34. doi: 10.1111/j.0906-6705.2005.00384.x.

Abstract

Ultraviolet B (UVB) induces both apoptosis and skin cancer. We found that human keratinocytes (KC) immortalized by Human Papillomavirus (HPV)16 E6/E7 were sensitized to UVB-induced apoptosis, possibly representing a transient regression-prone precancerous stage equivalent to actinic keratosis. To further examine which caspases are apical and essential, we utilized retroviral constructs expressing dominant-negative caspase-9 (caspase-9-DN) or Fas-associated protein with death domain (FADD)-DN as well as caspase inhibitor peptides. Caspase-9-DN and zLEHD-fmk both suppressed caspase-9, -3, and -8 activity after UVB exposure, as well as proteolytic processing of procaspase-3 into its active form, DNA fragmentation factor 45 cleavage, and internucleosomal DNA fragmentation. By contrast, stable expression of FADD-DN in HPV-immortalized KC did not inhibit UVB-induced activation of caspases-9, -3, and -8 nor downstream apoptotic events, although inhibition of caspase-8 with zIETD-fmk attenuated apoptosis. This study indicates that caspase-9 activation is upstream of caspases-3 and -8 and that UVB-induced apoptosis in HPV-immortalized human KC is death receptor (DR) independent and requires both caspase-9 upstream and caspase-8 downstream for maximal apoptosis. These studies further indicate that cell type as well as transformation state determine the sensitivity and mode of cell death (DR vs. mitochondrial apoptotic pathways) in response to UVB and explain the high regression rates of premalignant lesions.

Publication types

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

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Apoptosis / physiology*
  • Caspase 9
  • Caspases / metabolism*
  • Fas-Associated Death Domain Protein
  • Human papillomavirus 16
  • Humans
  • In Vitro Techniques
  • Keratinocytes / metabolism
  • Keratinocytes / radiation effects*
  • Male
  • Mitochondria / metabolism
  • Signal Transduction
  • Skin / metabolism
  • Ultraviolet Rays

Substances

  • Adaptor Proteins, Signal Transducing
  • FADD protein, human
  • Fas-Associated Death Domain Protein
  • CASP9 protein, human
  • Caspase 9
  • Caspases