Isolation of Melittin from Iranian Honey Bee Venom and Investigation of Its Effect on Proliferation of Cervical Cancer- HeLa Cell Line

Cervical cancer is the second prevalent cancer in developing countries and the sixth prevalent cancer in USA. Since conventional treatment methods are associated with detrimental side effects، searching for new drugs using natural ingredients is very important. Previous studies have shown that melittin (main component of honey bee venom) has anticancer properties along with the effect on cell membrane and activation of apoptosis. In this study، inhibitory effects of melittin on the viability and proliferation of cervical cancer cell line (HeLa) was investigated.

Melittin was purified from honeybee venom using reversed-phase HPLC method. Then، biological activity of melittin was examined by hemolytic activity analysis on the red blood cells. In order to investigate whether melittin inhibits proliferation of HeLa cell، MTT assay was performed. HeLa cells were plated in a 96-well plate and treated with serially diluted concentrations of melittin for 12 and 24 hours. The viability of the cells was measured via MTT assay at 540nm.

Melittin showed a strong hemolytic activity (HD50=0. 5 µg/ml) which can be reduced by FBS (HD50=2 µg/ml). Results of MTT assay indicated that melittin shows cytotoxic effect on cervical cancer cells with IC50 = 1. 2 ug/ml at 12h incubation period.

In this study، biological activity of melittin and inhibitory effect of FBS on hemolysis were determined via hemolytic activity analysis. MTT assay indicated that melittin induced cytotoxic effects in a dose dependent manner on cervical cancer cells and it also revealed dependence on incubation time as well.