مرکز تحقیقات بیولوژی کاربردی تکوین جانوری

Background: Malignant melanoma is a highly aggressive malignant melanocytic neoplasm which resists against the most conventional therapies. Sea cucumber as one of
marine organisms contains bioactive compounds such as polysaccharide, terpenoid
and other metabolites which have anti-cancer, anti-tumor, anti-inflammatory and
antioxidant properties. The present study was designed to investigate the anticancer
potential of saponin extracted from sea cucumber Holothuria leucospilata alone and
in combination with dacarbazine on B16F10 melanoma cell line.
Methods: The B16F10 cell line was treated with different concentrations of saponin (0,
4, 8, 12, 16, 20 μg/ml), dacarbazine (0, 1200, 1400, 1600, 1800, 2000 μg/ml) and coadministration of saponin-dacarbazine (1200 da+8 sp, 1200 da+4 sp) for 24 and 48 hr
and the cytotoxic effect was examined by MTT, DAPI, acridine orange/propodium iodide, flow cytometry and caspase colorimetric assay.
Results: The results exhibited that sea cucumber saponin, dacarbazine, and coadministration of saponin-dacarbazine inhibited the proliferation of melanoma cells
in a dose and time dependent manner with IC50 values of 10, 1400 and 4+1200 μg/ml,
respectively. Morphological observation of DAPI and acridine orange/propodium iodide staining documented typical characteristics of apoptotic cell death. Flow
cytometry assay indicated accumulation of IC50 treated cells in sub-G1 peak. Additionally, saponin extracted induced intrinsic apoptosis via up-regulation of caspase-3
and caspase-9.
Conclusion: These results revealed that the saponin extracted from sea cucumber as a
natural anti-cancer compound may be a new treatment modality for metastatic
melanoma and the application of sea cucumber saponin in combination with
dacarbazine demonstrated the strongest anti-cancer activity as compared with the
drug alone.