Cyclooxygenase-2 expression is up-regulated in obstructed human ureter

Stephen Y. Nakada, Travis Jerde, Lynn M. Jacobson, Ricardo Saban, Dale E. Bjorling, Debra A. Hullett

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Purpose: Prostanoids produce significant effects on ureteral function and are synthesized by cyclooxygenase (COX) enzymes. COX is found in the 2 isoforms COX-1 (a constitutive form) and COX-2 (an inducible form). Due to the side effects associated with COX-1 inhibition there is great interest in selective COX-2 inhibition. We determined if COX-2 messenger (m)RNA and protein expression are regulated during ureteral obstruction. Materials and Methods: mRNA analysis was performed using excess ureteral segments from 6 patients undergoing reconstructive procedures for chronic ureteral obstruction and 8 (normal ureter) undergoing donor nephrectomy after providing informed consent. All ureteral segments were snap frozen in liquid nitrogen and stored at -70C. RNA was isolated from the segments using phenol extraction and complementary DNA was synthesized by reverse transcription with murine leukemia virus reverse transcriptase (Promega Corp., Madison, Wisconsin). Semiquantitative reverse transcriptase-polymerase chain reaction was performed using specific COX-2 gene primers with ribosomal S26 primers serving as the housekeeping gene. The polymerase chain reaction product was quantified by agarose gel electrophoresis and phospho-imaging. The ratio of COX-2-to-S26 mRNA was compared. Additional segments were homogenized and total protein was extracted, separated via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. These membranes were Western blotted for COX-2 and glyceraldehyde-3-phosphate dehydrogenase (housekeeping protein) with specific primary and secondary antibodies. Results: The mean ratio of COX-2-to-S26 mRNA plus or minus standard error at 20, 22 and 24 cycles of amplification was 0.22 ± 0.04 in the 8 normal ureters compared with 1.01 ± 0.21 in the 6 obstructed ureters (unpaired Student's t test p = 0.004). Similarly the mean ratio of COX-2-to-glyceraldehyde-3-phosphate dehydrogenase protein on immunoblotting was 0.15 ± 0.02 in the 8 normal ureters compared with 0.59 ± 0.10 in the 6 obstructed ureters (p = 0.003) Conclusions: These data indicate that COX-2 mRNA and protein levels are up-regulated in chronically obstructed human ureters. Using selective COX-2 inhibitors may be useful for treating prostanoid induced effects associated with ureteral obstruction.

Original languageEnglish (US)
Pages (from-to)1226-1229
Number of pages4
JournalJournal of Urology
Volume168
Issue number3
StatePublished - Sep 2002
Externally publishedYes

Fingerprint

Cyclooxygenase 2
Ureter
Ureteral Obstruction
Messenger RNA
Cyclooxygenase 1
Glyceraldehyde-3-Phosphate Dehydrogenases
Prostaglandin-Endoperoxide Synthases
Proteins
Prostaglandins
Housekeeping
Murine Leukemia Viruses
Collodion
Membranes
Agar Gel Electrophoresis
Cyclooxygenase 2 Inhibitors
RNA-Directed DNA Polymerase
Essential Genes
Phenol
Reverse Transcriptase Polymerase Chain Reaction
Nephrectomy

Keywords

  • Cyclooxygenase inhibitors
  • Proteins
  • RNA, messenger
  • Ureter
  • Ureteral obstruction

ASJC Scopus subject areas

  • Urology

Cite this

Nakada, S. Y., Jerde, T., Jacobson, L. M., Saban, R., Bjorling, D. E., & Hullett, D. A. (2002). Cyclooxygenase-2 expression is up-regulated in obstructed human ureter. Journal of Urology, 168(3), 1226-1229.

Cyclooxygenase-2 expression is up-regulated in obstructed human ureter. / Nakada, Stephen Y.; Jerde, Travis; Jacobson, Lynn M.; Saban, Ricardo; Bjorling, Dale E.; Hullett, Debra A.

In: Journal of Urology, Vol. 168, No. 3, 09.2002, p. 1226-1229.

Research output: Contribution to journalArticle

Nakada, SY, Jerde, T, Jacobson, LM, Saban, R, Bjorling, DE & Hullett, DA 2002, 'Cyclooxygenase-2 expression is up-regulated in obstructed human ureter', Journal of Urology, vol. 168, no. 3, pp. 1226-1229.
Nakada SY, Jerde T, Jacobson LM, Saban R, Bjorling DE, Hullett DA. Cyclooxygenase-2 expression is up-regulated in obstructed human ureter. Journal of Urology. 2002 Sep;168(3):1226-1229.
Nakada, Stephen Y. ; Jerde, Travis ; Jacobson, Lynn M. ; Saban, Ricardo ; Bjorling, Dale E. ; Hullett, Debra A. / Cyclooxygenase-2 expression is up-regulated in obstructed human ureter. In: Journal of Urology. 2002 ; Vol. 168, No. 3. pp. 1226-1229.
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AU - Jerde, Travis

AU - Jacobson, Lynn M.

AU - Saban, Ricardo

AU - Bjorling, Dale E.

AU - Hullett, Debra A.

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N2 - Purpose: Prostanoids produce significant effects on ureteral function and are synthesized by cyclooxygenase (COX) enzymes. COX is found in the 2 isoforms COX-1 (a constitutive form) and COX-2 (an inducible form). Due to the side effects associated with COX-1 inhibition there is great interest in selective COX-2 inhibition. We determined if COX-2 messenger (m)RNA and protein expression are regulated during ureteral obstruction. Materials and Methods: mRNA analysis was performed using excess ureteral segments from 6 patients undergoing reconstructive procedures for chronic ureteral obstruction and 8 (normal ureter) undergoing donor nephrectomy after providing informed consent. All ureteral segments were snap frozen in liquid nitrogen and stored at -70C. RNA was isolated from the segments using phenol extraction and complementary DNA was synthesized by reverse transcription with murine leukemia virus reverse transcriptase (Promega Corp., Madison, Wisconsin). Semiquantitative reverse transcriptase-polymerase chain reaction was performed using specific COX-2 gene primers with ribosomal S26 primers serving as the housekeeping gene. The polymerase chain reaction product was quantified by agarose gel electrophoresis and phospho-imaging. The ratio of COX-2-to-S26 mRNA was compared. Additional segments were homogenized and total protein was extracted, separated via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. These membranes were Western blotted for COX-2 and glyceraldehyde-3-phosphate dehydrogenase (housekeeping protein) with specific primary and secondary antibodies. Results: The mean ratio of COX-2-to-S26 mRNA plus or minus standard error at 20, 22 and 24 cycles of amplification was 0.22 ± 0.04 in the 8 normal ureters compared with 1.01 ± 0.21 in the 6 obstructed ureters (unpaired Student's t test p = 0.004). Similarly the mean ratio of COX-2-to-glyceraldehyde-3-phosphate dehydrogenase protein on immunoblotting was 0.15 ± 0.02 in the 8 normal ureters compared with 0.59 ± 0.10 in the 6 obstructed ureters (p = 0.003) Conclusions: These data indicate that COX-2 mRNA and protein levels are up-regulated in chronically obstructed human ureters. Using selective COX-2 inhibitors may be useful for treating prostanoid induced effects associated with ureteral obstruction.

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