Down regulation of u-Pa by a nutrient mixture In hemangioma (eoma) cells by inducing Caspase-dependent apoptosis

Aim: To examine, whether the antitumor effects of a specific nutrient mixture are due to induction of apoptosis by inhibition of u-PA.

Materials and Methods: A nutrient mixture containing lysine, proline, ascorbic acid, and green tea extract which has showed anticancer activity against a number of cancer cell lines was used as an experimental composition. EOMA cells were grown in appropriate media with antibiotics in 24- well tissue culture plates. At near confluence, the cells were treated with nutrition mixture at 10, 100, 1000 μg/ml in triplicate.Analysis of u-PA activity was carried out by fibrin zymography. Morphological changes and caspase activation associated with apoptosis induction was checked by H&E staining and Live Green caspase assay, respectively. Apoptosis inducing anticancer drug camptothecin (10 μM) was used as positive control. Results: The nutrition mixture exhibited dose response toxicity with maximum toxicity 55% (p < 0.001) at 1000 μg/ml. EOMA cells expressed u-PA, which was inhibited by nutrition mixture in a dose-dependent manner. The caspase analysis revealed a dose dependent increase in apoptosis of EOMA hemangioma cells, with an increasing apoptosis observed at 100 μ g/ml, and maximum at 1000 μ g/ml. Cells treated with nutrition mixture showed significantly more apoptotic changes than the control or camptothecin-treated cells. Conclusion: These results suggest NM as a potential therapeutic agent as it specifically targets and induces apoptosis in hemangioma cells.

Key words: nutrient mixture, urokinase plasminogen activators, hemangioma, EOMA, caspase, apoptosis.