title: Adenovirus-mediated expression of cyclooxygenase-2 antisense reverse abnormal genetic profile of human adhesion fibroblasts.
abstract: OBJECTIVE
  To determine the effects of blocking the translation of cyclooxygenase-2 (COX-2) mRNA on the mRNA levels of type I collagen, type III collagen, fibronectin, and transforming growth factor-beta (TGF-beta1) in fibroblasts obtained from normal peritoneal and adhesion tissues.
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  University medical center.
  
  
  METHODS
  Fibroblasts established from peritoneal and adhesion tissue of the same patients.
  
  
  METHODS
  Adenovirus with COX-2 treatment of the primary cultured fibroblasts.
  
  
  METHODS
  Fibroblasts of normal peritoneal and adhesion tissues were isolated from the same patients. Adhesion and normal peritoneal fibroblasts were transfected with an adenovirus COX-2 mRNA in sense or antisense orientation. RNA was extracted from each treatment and subjected to real time reverse transcriptase-polymerase chain reaction to quantify change in mRNA levels of type I collagen, type III collagen, fibronectin, and TGF-beta1.
  
  
  RESULTS
  Type I collagen, type III collagen, fibronectin, and TGF-beta1 mRNAs were present in both normal peritoneal and adhesion fibroblasts with significantly higher levels in adhesion fibroblasts. Normal fibroblasts transfected with the COX-2 sense virus exhibited an elevated mRNA level in those molecules that reached the mRNA levels seen for adhesion fibroblasts. There were no effects seen on the mRNA levels in those molecules when adhesion fibroblasts were transfected with COX-2 sense virus. Normal fibroblasts transfected with COX-2 antisense virus exhibited no difference in mRNA levels in those molecules as compared with untransfected controls. In contrast, adhesion fibroblasts transfected with COX-2 antisense exhibited markedly reduced mRNA levels in those molecules to levels that were seen in normal peritoneal fibroblasts.
  
  
  CONCLUSIONS
  Our data suggest that inhibition of COX-2 may reduce the development of postoperative adhesions by preventing the formation of the adhesion phenotype.
authors.affiliation: Department of Obstetrics and Gynecology, C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan 48201, USA. gsaed@med.wayne.edu
authors.forename: Ghassan M
authors.initials: GM
authors.lastname: Saed
authors.forename: Ayman
authors.initials: A
authors.lastname: Al-Hendy
authors.forename: Salama A
authors.initials: SA
authors.lastname: Salama
authors.forename: Michael P
authors.initials: MP
authors.lastname: Diamond
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024061:Collagen Type III / Q000235:genetics / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000235:genetics* / Q000378:metabolism; D016373:DNA, Antisense / Q000235:genetics*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D010529:Peritoneal Cavity / Q000166:cytology; D011183:Postoperative Complications / Q000517:prevention & control; D012333:RNA, Messenger / Q000378:metabolism; D000267:Tissue Adhesions / Q000235:genetics*; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D024061:Collagen Type III; D016373:DNA, Antisense; D005353:Fibronectins; D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1; D051546:Cyclooxygenase 2