Assessment of cyclooxygenase inhibitors using in vitro assay systems
- 298 Downloads
Various inhibitors of cyclooxygenase are known to mediate cancer chemopreventive effects. We currently describe two in vitro assay systems for measuring cyclooxygenase activity. These assays can be used in combination and thereby provide a rapid, reliable, and economical approach that is applicable for large-scale evaluation of test samples. This approach employs peroxidase co-substrate oxidation and oxygen consumption assays. The former system, adapted to a 96-well plate format, detects inhibitors that function as a radical scavengers or interact with the enzyme directly. The latter system specifically monitors cyclooxygenase inhibitors that interact with the enzyme itself. Thus, the peroxidase co-substrate oxidation assay serves as a pre-screening method, whereas the oxygen consumption assay is used subsequently to investigate the mode of action mediated by samples which test positive.
Unable to display preview. Download preview PDF.
- 1.Boynton CS, Dick CF, Mayor GH (1988). NSAIDs: An overview. J Clin Pharmacol 28: 512–517.Google Scholar
- 2.Greenberg ER, Baron JA (1993). Prospects for preventing colorectal cancer death. J Natl Cancer Inst 85: 1182–1184.Google Scholar
- 3.Kune GA, Kune S, Watson LF (1988). Colorectal cancer risk, chronic illnesses, operations, and medications: Case control results from the Melbourne Colorectal Cancer Study. Cancer Res 48: 4399–4404.Google Scholar
- 4.Rosenberg L, Palmer JR, Zauber AG, Warshauer ME, Stolley PD, Shapiro S (1991). A hypothesis: Nonsteroidal antiinflammatory drugs reduce the incidence of large bowel cancer. J Natl Cancer Inst 83: 355–358.Google Scholar
- 5.Van der Ouderaa FJG, Buytenhek M (1982). Purification of PGH synthase from sheep vesicular glands. Meth Enzymol 86: 60–68.Google Scholar
- 6.Kulmacz RJ, Lands EM (1987). Cyclooxygenase: measurement, purification and properties. In: Benedetto C, McDonald-Gibson RG, Nigam S, Slater TF (eds), Prostaglandins and related substances: A practical approach. Oxford: IRL Press, Chap 12, pp 209–222.Google Scholar
- 7.Sano H, Hla T, Maier JAM, et al. (1992). In vivo cyclooxygenase expression in synovial tissues of patients with rheumatoid arthritis and osteoarthritis and rats with adjuvant and streptococcal cell wall arthritis. J Clin Invest 89: 97–108.Google Scholar
- 8.Kujubu DA, Herschman HR (1992). Dexamethasone inhibits mitogen induction of the TIS10 prostaglandin synthase/cyclooxygenase gene. J Biol Chem 267: 7991–7994.Google Scholar
- 9.Futaki N, Takahashi S, Yokoyama M, Arai I, Higuchi S, Otomo S (1994). NS-398, a new anti-inflammatory agent, selectively inhibits prostaglandin G/H synthase/cyclooxygenase (COX-2) activity in vitro. Prostaglandins 47: 55–59.Google Scholar
- 10.Gierse JK, Hauser SD, Creely DP, Koboldt C, Rangwala SH, Isakson PC, Seibert K (1995). Expression and selective inhibition of the constitutive and inducible forms of human cyclooxygenase. Biochem J 305: 479–484.Google Scholar
- 11.Jang M, Cai L, Udeani GO, Slowing K, Thomas CF, Beecher CWW, Fong HHS, Farnsworth NR, Kinghorn AD, Mehta RG, Moon RC, Pezzuto JM (1997). Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275: 218–220.Google Scholar