عزل ميليتين من سم النحل وتقييم تأثيره على انتشار خلايا سرطان المعدةIsolating Melittin from Bee Venom and Evaluating its Effect on Proliferation of Gastric Cancer Cells
عزل ميليتين من سم النحل وتقييم تأثيره على انتشار خلايا سرطان المعدة
Isolating Melittin from Bee Venom and
Evaluating its Effect on Proliferation of
Gastric Cancer CellsIsolating Melittin from Bee Venom and
Amir Mahmoodzadeh , Ali Morady1, Hannaneh Zarrinnahad
Kamran Pooshang Bagheri , Pooria Ghasemi-Dehkordi , Mahdi Mahdavi
Delavar Shahbazzadeh , Hamidreza Shahmorady
Background: Gastric cancer (GC) is one of the most common cancers worldwide and
in Iran. As conventional therapies such as surgery and chemotherapy are invasive with
adverse effects, current studies are important as they are conducted to find natural compounds
with few adverse effects. In this study, melittin with 26 amino acids was isolated
and purified from bee venom and its effect on the viability and proliferation of gastric
cancer cells was investigated.
Methods: At first, melittin was purified from honeybee venom by a reversed-phase high
performance liquid chromatography (RP- HPLC) and using C18 column. The biologic
activity of melittin was evaluated by hemolytic test on red blood cells to melittin standard.
To investigate the effect of melittin on viability and proliferation of AGS human
gastric adenocarcinoma cells, the related cells were cultured in a 96-well plate and treated
with serially diluted concentrations of melittin for 6 and 12 hours and their mortality was
determined by MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]
colorimetric method at 540 nm wavelengths.
Results: The obtained chromatogram from RP-HPLC showed that melittin comprised
50% of the studied bee venom. SDS-PAGE analysis of melittin fraction confirmed purity
of isolated melittin. Hemolytic assay showed that the extracted melittin indicates a strong
hemolytic activity (HD50=0.55μg/ml). MTT assay showed that melittin strongly inhibits
proliferation of gastric cancer cells at concentrations more than 2μg/ml. This inhibitory
effect depends on melittin concentration and time.
Conclusion: The results of this study showed that melittin is a strong inhibitor of proliferation
of the gastric cancer cells. Also, it was observed that this inhibitory effect is
increased with increasing concentrations of melittin and incubation time.
Keywords: Melittin, Gastric Cancer, Reverse-Phase Chromatography.