title: Antitumor effect of reduction of 150-kDa oxygen-regulated protein expression on human prostate cancer cells.
abstract: BACKGROUND
  The 150-kDa oxygen-regulated protein ORP150, a new member of the heat shock protein family that functions as a molecular chaperone in the endoplasmic reticulum, was found to increase in infiltrating cancer cells. Since enhancement of ORP150 expression and the presence of vascular endothelial growth factor (VEGF) in human prostate cancer glands were immunohistochemically demonstrated, we examined whether transduced antisense ORP150 cDNA can reduce angiogenicity and tumorigenicity through suppression of VEGF secretion.
  
  
  METHODS
  Human prostate cancer specimens were immunohistochemically stained with fluorescein isothiocyanate (FITC) for ORP150 or vascular endothelial growth factor (VEGF). An adenovirus vector (Ad) carrying antisense ORP150 cDNA (AdCA-Antisense ORP150) was constructed and infected to prostate cancer DU145 cells. Expression of ORP150 in the cells was analyzed with western blotting and secretion of VEGF into the supernatant with an enzyme-linked immunoabsorbent assay (ELISA). Angiogenicity was evaluated by chorioallantoic membrane (CAM) assay. A nude mouse xenograft model was used to examine tumorigenicity.
  
  
  RESULTS
  Immunohistochemical study proved that the expression of ORP150 and VEGF was enhanced in the cytoplasm of prostate cancer cells. The Ad showed 100% transduction efficiency and minimum cytotoxicity when the cells were infected at a multiplicity of infection (MOI) of 20 for 24 h. Expression of ORP150 was substantially reduced by the antisense treatment. Secretion of VEGF into the culture supernatant was reduced to 30%. Consequently, the CAM assay showed relatively low angiogenicity, while marked suppression of tumor formation was observed in the xenograft model.
  
  
  CONCLUSIONS
  Adenoviral-mediated antisense ORP150 cDNA transfer is well worth considering as an option for prostate cancer gene therapy.
authors.affiliation: Department of Urology, Kanazawa University School of Medicine, Kanazawa, Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Miyagi
authors.forename: Osamu
authors.initials: O
authors.lastname: Hori
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Koshida
authors.forename: Masayuki
authors.initials: M
authors.lastname: Egawa
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kato
authors.forename: Yasuhide
authors.initials: Y
authors.lastname: Kitagawa
authors.forename: Kentaro
authors.initials: K
authors.lastname: Ozawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ogawa
authors.forename: Mikio
authors.initials: M
authors.lastname: Namiki
mesh_terms: D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D016373:DNA, Antisense / Q000235:genetics; D016228:Endothelial Growth Factors / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D018840:HSP70 Heat-Shock Proteins; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D008222:Lymphokines / Q000235:genetics; D008297:Male; D051379:Mice; D008819:Mice, Nude; D023421:Models, Animal; D018832:Molecular Chaperones / Q000235:genetics / Q000494:pharmacology*; D009389:Neovascularization, Pathologic; D011471:Prostatic Neoplasms / Q000098:blood supply / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D011506:Proteins / Q000235:genetics / Q000494:pharmacology*; D014161:Transduction, Genetic / Q000379:methods; D014407:Tumor Cells, Cultured / Q000187:drug effects*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D016373:DNA, Antisense; D016228:Endothelial Growth Factors; D018840:HSP70 Heat-Shock Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D018832:Molecular Chaperones; D011506:Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; C050087:oxygen-regulated proteins