Cytotoxic effect of sea anemone pore-forming toxin on K۵۶۲ chronic myeloid leukemia cells

Chronic myelogenous leukemia (CML) is one of prevalent cancer worldwide. In spite of various designed drugs, chemoresistance remains the main obstacle in cancer cure. Therefore, developing novel strategy for treatment of CML is an urgent need. Fragaceatoxin C (FraC) is novel protein toxin from a sea anemone called actinia fragacea with great impacts against cells by pore formation and disturbing cell membrane integrity. The aim of this study was evaluation of FraC toxin toxicity against K۵۶۲. The bacteria cells harboring expression vector of FraC were induced by IPTG and purified by Ni۲+-NTA sepharose affinity chromatography. Then, purified toxin activity was evaluated using RBC hemolytic test. Eventually, evaluation of FraC cytotoxicity and apoptosis were performed using MTT and flow cytometery assays, respectively. Our results revealed that FraC toxin decreased K۵۶۲ cells viability in a dose- and time-dependent manner with a whole destroy of cancer cells at ۳۵.۰۰ µg mL-۱ after ۷۲ hr. Furthermore, flow cytometery analysis indicated that FraC toxin enhanced necrosis along with apoptosis in K۵۶۲ cells in a dose dependent manner. We speculated that FraC toxin could be considered as a novel candidate for cancer cell researches and treatments provided that it should be turned into a specific agent by engineering and directing to cancer cell membrane.

Keywords:

Chronic myelogenous leukemia , Fragaceatoxin C , Necrosis , Pore-forming toxin