BREAST CANCER CELLS INDUCED APOPTOSIS BY T47D 60 kDa PROTEIN OF RABBIT INK SEA (Aplysia dactylomela, Rang 1828)
ABSTRACT: Sea hares (Aplysia dactylomela Rang, 1828) is marine Mollusks that secretes poisonous purple inks as resistance against predators. Ink from Aplysia has been reported contains 60 kDA protein which has potency as antibacterial, antioxidant, and anticancer. Apoptosis is one of cancer inhibition mechanism. The aim of this research was to determine the potency of sea hare ink (Aplysia dactylomela Rang, 1828) to induce apoptosis on T47D cell line for breast cancer modell. Aplysia has been identified based on morphology. Ink sampling was done by massaging the ink gland of the sea hares. After that, the ink was purified by SDS-PAGE to obtain 60 kDa protein, then the concentration was measured by Bradford assay. There were 4 groups for apoptosis test, which were 60 kDa protein with concentration of 3, 7, and 15µg/mL. Positive control: doxorubicin with concentration of 3, 7, and 15µg/ml. Solvent control use PBS and cell control as negative one. Double staining with ethidium bromide-acridine orange method was used to stain the cell. Apoptotic cell was viewed under fluorescence microscop.Viable cell was stained green, the apoptosis one was stained fluorescently orange. The number of apoptotic cell was measured in percentage. The results have shown that the species has been used is Aplysia dactylomela. The apoptosis test result shows that 60 kDa protein treatment induces apoptosis in breast cancer T47D cell culture. The percentages of apoptosis has occurred at 65.33; 85.33; and 96.67% respectively for protein concentration of 3, 7, and 15 µg/ml. The conclusion is that higher concentrations induce more apoptosis.
