CYTOTOXIC and GELATINOLYTIC ACTIVITIES of Mesobuthus Gibbosus (Brullé, 1832) VENOM

The crude venom of Mesobuthus gibbosus which is widely distributed in the Mediterranean region of Anatolia were investigated for its cytotoxic and gelatinolytic activities in vitro. Cytotoxic effects of the crude venom were examined in three different cell lines by MTT assay. 5RP7 is H-ras activated mouse fibroblasts, CO25 is N-ras activated mouse myoblasts and A549 is human adenocarcinomas cells. For the gelatinolytic activity, lyophilized crude venom was solubilized in bidistilled water, then run on a SDS/PAGE-gelatin. The gel is then stained with R-250 Commassie blue and gelatinolytic activity was detected as colorless bands. Effects of the venom on the morphology of 5RP7 cells were also examined by Giemsa staining. The number of viable 5RP7 cells was significantly decreased after incubation with the venom for 48 hours and IC50 was found 200 g /ml, whereas any effect of the venom on the viability of CO25 was not observed. The number of viable A549 cells was inhibited % 40 at the lowest dose (10 g /ml) after 24 h incubation and the rate of this inhibition was elevated up to % 60 dose-dependent manners. Mesobuthus gibbosus crude venom and components showed a significant gelatinolytic activity. The 5RP7 cells treated with 200 g /ml and 400 g/ml of venom were smaller rounded in shape; especially nuclei were stained more densely than their normal counterparts. Cytoplasmic blebs around the cells and also vacuole formation were observed. We suggest that this study will lead to identification of active peptides in M. gibbosus venom and following its use in cancer treatment.