Neovascularization is a major challenge in different ophthalmologic pathologies e.g. AMD, PDR, neovascular glaucoma. The current treatment strategies are surgery, photocoagulation and antiproliferative medical therapy. To improve the treatment of these patients further antiproliferative agents are warranted. Therefore, we treated human microvascular endothelial cells (HMVEC) with genistein (inhibitor of the protein tyrosine kinase and topoisomerase II) to elucidate its antiproliferative effects.

Methods: HMVEC from adult dermis (TEBU, D-60596 Frankfurt a/M) were cultured in wells until a confluent cell layer was achieved. Cultures were then incubated for 18 hours in Medium 131 with different genistein concentrations (12,5µg/ml, 25µg/ml, 50µg/ml and 100µg/ml). Medium + DMSO (10µl/ml) served as control. The cell proliferation was measured with the reagent WST-1 (Boehringer-Mannheim, Germany). The amount of protein p53 was measured using a p53 pan ELISA kit from Boehringer-Mannheim, Germany.

Results: The cell number was reduced significantly at concentrations of ³ 25µg/ml genistein. The level of p53 (control) decreased in correlation with the reduced cell numbers, stabilized at concentrations of >= 25µg/ml.

Conclusions: A significant reduction of HMVEC proliferation in vitro was found at concentrations of >= 25µg/ml genistein. P53 is detectable in normal HMVECs. The constant level of p53, while the cell number decreased, is suggesting that genistein induces apoptosis in HMVEC.

Dep. of Ophthalmology, University of Cologne; D-50924 Cologne

Supported in part by Retinovit Foundation

Back