Oxidative stress mediates cell surface expression of SS-A/Ro antigen on keratinocytes

Free Radic Biol Med. 2002 May 15;32(10):1006-16. doi: 10.1016/s0891-5849(02)00797-9.

Abstract

Exposure to ultraviolet radiation exacerbates the skin lesions of autoimmune diseases, and is known to induce cell surface expression of SS-A/Ro antigen on keratinocytes in vitro. Following up on recent reports on ultraviolet-B (UVB)-induced oxidative stress, we examined the role of oxidative stress in the surface expression of SS-A/Ro antigen on human keratinocytes. First, the exclusive induction by UVB irradiation of the 52-kDa protein (Ro52) but not of the 60-kDa protein (Ro60) of SS-A/Ro antigen was demonstrated by means of indirect immunofluorescence. The surface expression of Ro52 induced by UVB irradiation was concentration-dependently inhibited by N-acetyl-L-cysteine, an antioxidant. Furthermore, surface expression of Ro52 was similarly induced by diamide, a chemical oxidant. We next used Hoechst 33342 staining and the TUNEL assay to demonstrate that a low dose (20 mJ/cm(2)) of UVB did not induce apoptosis but induced the surface expression of Ro52. Moreover, zVAD-fmk, a pan-caspase inhibitor, did not inhibit UVB-induced surface expression of Ro52 even at a high dose (200 mJ/cm(2)) of UVB, which was sufficient to induce apoptosis in keratinocytes in the absence of zVAD-fmk. Taken together, we concluded that UVB-induced surface expression of Ro52 on keratinocytes is mediated by oxidative stress through a pathway other than apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine / pharmacology
  • Amino Acid Chloromethyl Ketones / pharmacology
  • Antigens, Surface / metabolism*
  • Apoptosis
  • Autoantigens / metabolism*
  • Autoimmune Diseases / etiology*
  • Benzimidazoles
  • Blotting, Western
  • Caspase Inhibitors
  • Cells, Cultured
  • Diamide / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Fluorescent Dyes
  • Free Radical Scavengers
  • Humans
  • In Situ Nick-End Labeling
  • Infant, Newborn
  • Keratinocytes / cytology
  • Keratinocytes / metabolism*
  • Keratinocytes / radiation effects
  • Oxidative Stress*
  • RNA, Small Cytoplasmic*
  • Ribonucleoproteins / antagonists & inhibitors
  • Ribonucleoproteins / metabolism*
  • Ultraviolet Rays

Substances

  • Amino Acid Chloromethyl Ketones
  • Antigens, Surface
  • Autoantigens
  • Benzimidazoles
  • Caspase Inhibitors
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • Free Radical Scavengers
  • RNA, Small Cytoplasmic
  • RO60 protein, human
  • Ribonucleoproteins
  • SS-A antigen
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • Diamide
  • bisbenzimide ethoxide trihydrochloride
  • Acetylcysteine