P-27: Trypsin Inhibitor Purified from Cucumis Melo Seeds: A Possible Therapeutic Perspective for Tumor and Angiogenesis Related Disease


Mansouri K *, Parvaneh Sh , Mostafaie A , Mahnam A , Mohammadi-Motlagh HR ,

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Objective: Angiogenesis, the formation of new blood vessels, is a key process in cancer development and metastasis. In this study, anti-angiogenesis and anti-proliferative potential of Trypsin inhibitor purified from cucumis melo seeds (TIPCM) has been investigated using human umbilical vein endothelial cells (HUVEC). Materials and Methods: Anti-angiogenesis effect of Trypsin inhibitor purified from cucumis melo seeds (TIPCM) on cell migration and tubulogenesis in microcarrier-based fibrin gel was assayed microscopically. HUVEC were isolated from human umbilical vein of newborn babies by collagenase digestion method. The isolated cells were grown in MCDB-131 media supplemented with 20% FBS. HUVEC at passage 2 to 3 were used in this experiment. For in vitro system of angiogenesis, the cells were cultured in a standard microcarrier-based model in fibrin gel. The cultured cells were treated with (TIPCM) at 0, 5, 10, 20, 40 and 60 μg/ml of (TIPCM) in triplicate at each dose for migration and tubulogenesis.In the days 5–7, angiogenesis was clearly visible as spike-like protrusions from the particles. Results: Electrophoresis analysis of purified protein shows a single band with a molecular mass of about 3 KDa .Effect of the (TIPCM) at different doses on cell migration and angiogenesis were shown .angiogenesis of the endothelial cells treated with purified (TIPCM) at 5, 10, 20, 40 and 60 μg/ml respectively. In these conditions,migration and tubulogenesis of the endothelial cells through the fibrin matrix were inhibited completely. Conclusion: All the tested doses inhibited blood vessel formation but had no toxic effect on endothelial cell. Because of these properties, (TIPCM)can be used as a specific angiogenesis inhibitor. Combination of this protein with cytotoxic agents may be used in pathological process such as tumor metastasis