Fractination of anticancer protein from a marine fungus Xylaria psidii KT30 and their cytotoxicity against HeLa cell line.

Abstract

Cervical cancer is the most common death cause of cancer in Indonesia after human breast cancer. One of the efforts of cancer treatment is the utilization of natural product. Some microorganism have potential as anticancer such as endophytic fungi. Endophytic fungi from the marine habitats can be isolated from seaweeds, seagrasses, sponges, and mangroves. Xylaria psidii KT30 is fungus from seaweed that can produce anticancer protein. This study aimed to obtain protein fraction from Xylaria psidii KT30 and determine their toxicity against Chang and HeLa cells. The research was carried out in 3 stages. Phase 1 Xylaria psidii KT30 was cultivated in Potato Dextrose Agar (PDA) and Potato Dextrose Broth (PDB) medium, the metabolites was extracted using 90% of ammonium sulfate. Phase 2 the toxicity was determined by Brine Shrimp Lethality Test (BSLT). Phase 3 was the final stage, the fractionation process was conducted using DEAE Sephadex A-50 column chromatography and cytotoxic assay using Chang (ATCC CCL 13) and HeLa (ATCC CCL 2) cells. Xylaria psidii KT30 yielded 0,090% of extracellular protein. The results revealed that LC50 of protein extract was 104,95 ppm. Fractionation of protein produced five protein fraction groups comprised of fraction I (2-12), fraction II (13-49), fraction III (50-61), fraction IV (62-72), and fraction V (73-100). Based on the results of SDS-PAGE, the molecular weight of selected fractions were 23,99; 32,88 and 37,30 kDa. Cytotoxicity assay results in Chang cells showed that the crude extract and fractions of selected proteins (F3.1, F3.2, and F4) was not active against normal cells with inhibition less than 50%, while in IC50 value of HeLa cell protein fractions F3.1, F3.2 and F4 against cervical cancer cells (HeLa) in this study could not be determined because in highest concentration (1080 mg/mL) did not produce % inhibition more than 50% of the HeLa cells. The results revealed that IC50 of the crude extract proteins of marine fungus was 69,89 μg/mL.