Inducing Apoptosis of Cancer Cells Using Sea Pen Virgularia gustaviana Extract Which is Comparable to Cembrane Diterpene Sarcophine

Marine Soft corals have frequently been studied in recent years because of their specific chemical compounds in tissue engineering and regenerative medicine. The aim of this study was to investigate anti-cancer property of extracted compound from Virgularia gustaviana and their effect on inducing of apoptosis. The extraction process was carried out with ethyl acetate for 5 days and the extract was separated by silica-gel column chromatography. The column was washed with n-hexane-ethyl acetate solvent at ratio of 10:0 to 0:10. Thin layer chromatography (TLC), High performance thin layer chromatography (HPTLC), high performance liquid chromatography (HPLC), and 13C NMR spectroscopy were used for qualitative identification of compounds. The viability of HeLa and MDA-MB-231 cancer cells was investigated using MTT assay at the concentrations of 25, 50, and 100 µL/mL of extracted compounds. Immunocytochemistry and Western Blot analyses were used to evaluate expression of apoptotic markers caspase-3 and caspase-8 in cancer cells after treating with effective fractions (based on viability of cancer cells) and the results were compared with Sarcophine. From ten isolated fractions (A-J), Retention time and Retention Factors (Rf) of fractions G, I, and J were the same as Sarcophine. Fraction G, I, and J dose-dependently decreased cancer cell viability compared to control group and Sarcophine. Treatment of cancer cells with the latest fraction increased expression of caspase-3 and caspase-8 demonstrating induction of apoptosis as possible mechanism of action. According to the results, the compounds extracted from Virgularia gustaviana inhibit the growth of cancer cells by inducing of apoptosis pathway; an effect which needs to be further investigated in the future studies.