In-vitro Cytotoxic Anticancer Effects of Honeybee Venom Fractions on Different Cell Lines

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Montaha A. Al-Safar
Hamid Naji Obied
Rana A. Ghaleb
Ali S. Kashkol


Background: Honey bee venom (HBV) has been used from ancient history in traditional medicine, but recently with the progression of medical and biological science, some HBV fractions were found to had specific therapeutic use. Honey bee venom fractions such as melittin were noticed to have had in-vivo and in-vitro anti-tumor effects. Honey bee venom has about twenty different biological fractions. Melittin was the dominant fraction extracted from crude HBV.
The aim is to study the in-vitro anti-cancer effect of one HBV fractions extracted by reverse-phase high-performance liquid chromatography (RP-.HPLC) against four cancer and one none tumorigenic normal cell line.HBV was collected by using the electric collector and dried by lyophilization and separated by RP-HPLC into sixteen different weights yield uncharacterized fractions. For detection of the anti-cancer effect, all the 16 HPLC peaks were tested separately on four cancer cell lines and one none tumorigenic cell line.
Materials and methods: Four cancer cell lines, B16, MCF-7, Hep-G2, HCT-116, and MDCK normal canine kidney cell line was cultured and maintained on RBMI-1640 media with 10% fetal bovine serum. The HBV 16 fractions was tested on the above cell lines separately by using MTT cytotoxicity assay protocol. The viability percent was obtained versus five concentrations of HBV (50, 25, 12.5, 6.25, 3.125 Ug). The half-maximal inhibitory concentration (IC50) was calculated and plotted using sigma plot software, it was found as 5.6 for B16, 21.2 for MCF-7.14 for Hep-G2, 21.4 for HCT116, and 45 for MDCK cell line subsequently.
Cell line subsequently from the result we found that only fraction number 8 (HBv-F8), has had cytotoxic effects on the cancer cell lines and the normal canine kidney cell line while the other fifteen HPLC fraction had no any cytotoxic effects.
Conclusion: We conclude that HBV-F8 has a cytotoxic effect on cancer and normal cell lines with different IC 50 for each cell line. The best cytotoxic effect was found on B16 melanoma cell line, and the least effect was found on HCT-116 cell line. At the same time, it is also toxic on MDCK normal cell line at 45.13 ug half-maximal inhibitory concentration.

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