Adenovirus-mediated expression of cyclooxygenase-2 antisense reverse abnormal genetic profile of human adhesion fibroblasts

Ghassan M. Saed, Ayman Al-Hendy, Salama A. Salama, Michael Peter Diamond

Research output: Contribution to journalArticle

7 Citations (Scopus)

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-β1) in fibroblasts obtained from normal peritoneal and adhesion tissues. Design: Prospective experimental study. Setting: University medical center. Patient(s): Fibroblasts established from peritoneal and adhesion tissue of the same patients. Intervention(s): Adenovirus with COX-2 treatment of the primary cultured fibroblasts. Main Outcome Measure(s): 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-β1. Result(s): Type I collagen, type III collagen, fibronectin, and TGF-β1 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. Conclusion(s): Our data suggest that inhibition of COX-2 may reduce the development of postoperative adhesions by preventing the formation of the adhesion phenotype.

Original languageEnglish (US)
Pages (from-to)1455-1460
Number of pages6
JournalFertility and sterility
Volume89
Issue number5 SUPPL.
DOIs
StatePublished - May 1 2008

Fingerprint

Reverse Genetics
Cyclooxygenase 2
Adenoviridae
Fibroblasts
Messenger RNA
Tissue Adhesions
Collagen Type III
Fibronectins
Collagen
Viruses
Reverse Transcriptase Polymerase Chain Reaction
Transforming Growth Factor beta
Real-Time Polymerase Chain Reaction

Keywords

  • Adenovirus
  • COX-2
  • fibroblasts
  • peritoneium
  • postoperative adhesions

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology

Cite this

Adenovirus-mediated expression of cyclooxygenase-2 antisense reverse abnormal genetic profile of human adhesion fibroblasts. / Saed, Ghassan M.; Al-Hendy, Ayman; Salama, Salama A.; Diamond, Michael Peter.

In: Fertility and sterility, Vol. 89, No. 5 SUPPL., 01.05.2008, p. 1455-1460.

Research output: Contribution to journalArticle

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AB - 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-β1) in fibroblasts obtained from normal peritoneal and adhesion tissues. Design: Prospective experimental study. Setting: University medical center. Patient(s): Fibroblasts established from peritoneal and adhesion tissue of the same patients. Intervention(s): Adenovirus with COX-2 treatment of the primary cultured fibroblasts. Main Outcome Measure(s): 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-β1. Result(s): Type I collagen, type III collagen, fibronectin, and TGF-β1 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. Conclusion(s): Our data suggest that inhibition of COX-2 may reduce the development of postoperative adhesions by preventing the formation of the adhesion phenotype.

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