title: Gene expression profiling of Ca2+-atpase inhibitor DTBHQ and antigen-stimulated RBL-2H3 mast cells.
abstract: OBJECTIVE
  Ca2+ signaling is critical for mast cell activation by antigen stimulation, and we previously described that the signaling can be mimicked by Ca2+-ATPase inhibitors. We therefore investigated the effect of the Ca2+-ATPase inhibitor and antigen stimulation on the gene expression profiles of RBL-2H3 mast cells.
  
  
  METHODS
  A Ca2+-ATPase inhibitor, 2,5-di(tert-butyl)-1,4-hydroquinone (DTBHQ), an antigen (dinitrophenylated BSA), a high-density oligonucleotide microarray (Affymetrix GeneChip) technique, and a well-characterized rat mast cell line RBL-2H3 were used.
  
  
  METHODS
  RBL-2H3 cells were activated for 3 h with 10 microM DTBHQ, which increases cytosolic Ca2+ concentration, or 10 microg/ml antigen, which cross-links IgE receptors, and the mRNA expression profiles (8,799 genes) were analyzed with GeneChip arrays (n = 3).
  
  
  METHODS
  Expression levels were measured by GeneChip, and the differences were tested by Welch's t-test and P-values less than 0.05 were considered statistically significant. Values are expressed as means +/- SEM.
  
  
  RESULTS
  The genes, including MCP-1, GADD45, Relaxin H1, CSF-1, c-jun-oncogene, Pyk-2, NKR-P2 and CREM, were significantly up-regulated by both DTBHQ and antigen stimuli, whereas the genes including interleukin (IL)-3, IL-4, IL-9, IL-13, GADD153, butyrate response factor, and Fas ligand, were up-regulated by DTBHQ alone. On the other hand, the expression of several genes, including GATA-1, were down-regulated by DTBHQ stimulation.
  
  
  CONCLUSIONS
  These results suggest 1) that DTBHQ seems to induce proinflammatory responses by stimulating the production of several cytokines through the expression of several transcription factors, 2) that the changes in gene expression profile induced by DTBHQ and by IgE receptor cross-linking in mast cells were almost the same, but many more stress-inducible genes like GADD 153 were up-regulated by the former.
authors.affiliation: Division of Biochemistry and Immunochemistry, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Nakamura
authors.forename: S
authors.initials: S
authors.lastname: Ishida
authors.forename: S
authors.initials: S
authors.lastname: Ozawa
authors.forename: Y
authors.initials: Y
authors.lastname: Saito
authors.forename: H
authors.initials: H
authors.lastname: Okunuki
authors.forename: R
authors.initials: R
authors.lastname: Teshima
authors.forename: J
authors.initials: J
authors.lastname: Sawada
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics*; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D015536:Down-Regulation / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006873:Hydroquinones / Q000494:pharmacology*; D007073:Immunoglobulin E / Q000494:pharmacology; D008407:Mast Cells / Q000187:drug effects / Q000201:enzymology*; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D000975:Antioxidants; D016207:Cytokines; D006873:Hydroquinones; C019359:2,5-di-tert-butylhydroquinone; D007073:Immunoglobulin E; D000252:Calcium-Transporting ATPases