Apoptotic Effect of Melittin Purified from Iranian Honey Bee Venom on Human Cervical Cancer HeLa Cell Line

Hannaneh Zarrinnahad,Delavar Shahbazzadeh,Amir Mahmoodzadeh,Monireh Parviz Hamidi,Kamran Pooshang Bagheri,Mehdi Mahdavi,Ali Moradi

doi:10.1007/s10989-017-9641-1

Abstract

Melittin, an amphipathic 26-residue peptide, is the main component of honey bee venom. Studies have been demonstrated that melittin has an inhibitory effect on proliferation of cancer cells. However, the precise mechanism of action is not completely understood. In the present study we have shown that purified melittin from Iranian honey bee venom shows anti-cancer effects on human cervical cancer cell line through induction of apoptosis. The venom was collected from Iranian honey bee (Apis mellifera meda) and melittin isolated using reversed phase HPLC. Biological activity of melittin was analyzed by hemolytic test on human red blood cells. In order to investigate whether melittin inhibits proliferation of cervical cancer cells, the viability of the melittin treated HeLa cell line was measured via MTT assay. Finally, cell death analysis was performed using Propidum iodide and Annexin V-FITC dual staining. The results showed that the half hemolytic concentration (HD50) induced by mellitin was 0.5 µg/ml in free FBS solution. IC50 obtained after 12 h at 1.8 µg/ml by MTT assay. According to flow cytometric analysis, melittin induced apoptosis at concentrations more than 1 µg/ml. These results suggest that melittin induces apoptotic cell death in cervical cancerous cells as observed by flow cytometric assay. It is concluded that melittin could be regarded as a potential candidate in future studies to discovery of new anticancer agents.

Highlights

IntroductionHoney Bee Venom (HBV) is a colorless liquid, with acidic entity (pH 4.5–5.5) (Kim et al 2007)
Honey Bee Venom (HBV) is a colorless liquid, with acidic entity (Kim et al 2007)
Results of MTT assay showed that melittin inhibited the proliferation of Hela cell line in a dose and time dependent manner (Fig. 6)

Summary

Introduction

Honey Bee Venom (HBV) is a colorless liquid, with acidic entity (pH 4.5–5.5) (Kim et al 2007). HBV has at least 28 different active compounds with a distinguished health benefits. Several studies identified biological properties of these compounds and the health benefits of each ingredient (Moon et al 2008). HBV is composed of various peptides including melittin, apamin, adolapin and Mast cell degranulationpeptide (MCD), enzymes such as phospholipase A2 and Evidences show that the activation of apoptosis process plays a critical role in treatment of cancers. Bee venom and melittin can induce apoptosis in several cancer cell lines such as human lung cancer cell line NCI-H1299(Jang et al 2003),. Melittin prevents liver cancer cell metastasis through inhibition of the Rac1-Dependent pathway (Rady et al 2017)

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