Visit our Expo - Redox and Inflammation signaling 2012

Session XVII : Cell signaling in health and disease Poster XVII, 2
2-Methoxyestradiol inhibits superoxide anion generation while enhaces superoxide
dismutase activity in swine granulosa cells
Giuseppina Basini, Sujen E. Santini, Francesca Grasselli
Dipartimento di Produzioni Animali, Biotecnologie Veterinarie, Qualità e Sicurezza
degli Alimenti, Sezione di Fisiologia Veterinaria, Università di Parma, 43100 Parma,
Italy. E-mail: basini@unipr.it
Angiogenesis and antiangiogenesis physiologically take place during ovarian follicular
growth and regression. Antiangiogenesis has been the subject of intense interest because of its
implications for restricting tumor growth. 2-Methoxyestradiol (2-ME) is an estradiol
metabolite with antiangiogenic and antitumor activity. It is formed by granulosa cell and is
present in the normal follicle at high concentrations. In this unique microenvironment, it may
regulate selected cell types via autocrine and/or paracrine action. Identification of reactive
oxygen specie role in the molecular pharmacology of 2-ME is an active area of research. To
this purpose we evaluated the effect of 2-ME on both superoxide anion (O2-) production
(WST-1 test, Roche, Mannheim, Germany) and on superoxide dismutase activity (SOD Assay
kit Dojndo Molecular Technologies, Japan) in swine granulosa cells. Cells were obtained
from follicles > 5 mm, seeded in 200 ul M199 in 96well plates and treated for 48 h with 1 uM
2-ME. O2- assay was performed on 104 cells/well and 20 ul WST-1 were added during the
last 4 h of incubation. SOD assay was performed on 2'105 cells/well; after treatment, cells
were lysed and assayed for enzyme activity. In both assays the absorbance was read at 450
nm against 620 nm. 2-ME inhibited (p