Registration number: 2020/39/O/NZ3/01760
Financing institution: National Science Centre
Project manager: prof. Jacek Kurzepa, MD, PhD
Project executor (PhD student): MSc Agata Małek
Place of implementation: Department of Medical Chemistry, Faculty of Medical Sciences, Medical University of Lublin
Budget: PLN 538,680
Implementation time: Oct 01, 2021 - Sep 30.2025
Among the many types of cancer, those affecting the nervous system that develop inside the skull, such as glioma, are of particular concern. One of the natural substances with potential anti-cancer properties is bee venom, also known as apitoxin. This compound, associated rather with unpleasant experiences related to stings, is a mixture of many biologically active ingredients that may have pharmaceutical applications. The first reports of the anti-cancer properties of bee venom appeared in the 1950s. Later observations reported a lower incidence of cancer among beekeepers, which was explained by more frequent stings compared to the rest of the population. It is now known that bee venom has anti-cancer properties for many types of cells, but the mechanism of its action seems to be both complex and not the same for different types of cancer. However, there is very little literature data on the effect of bee venom on glioma cells, despite the fact that apitoxin components demonstrate the ability to cross the blood-brain barrier, which effectively limits the penetration of many therapeutic substances into the nervous system. This project aims to investigate the anti-glioma properties of bee venom, both using glioma cell lines and examining the effect of the venom on glioma cells implanted into zebrafish embryos.
Bee venom (BV, also known as apitoxin) is synthesized and secreted by a venom gland present in the abdominal cavity of a worker bee or queen bee. It is a very complex mixture containing over 300 biologically active ingredients with a wide range of pharmacological properties. The vast majority of its dry mass is peptide compounds (melittin, apamin, mast cell degranulating peptide) and enzymes as phospholipase A2. The above figure on the left shows a chromatogram of BV. In the above figure, the highest peak (marked with an arrow) comes from melittin, which is the most abundant in bee venom. In our project, we fractionated the BV, i.e. divided it into parts, each containing different active substances. Then we assessed which of the obtained fractions showed anti-glioma effects. The results of research showing the effect of the obtained BV fractions on glioblastoma cell cultures were presented in the original publication "Anticancer Activity of Melittin...".
Honey bee venom in its composition contains many biologically active peptides and enzymes that are effective in the fight against diseases of various etiologies. The history of the use of bee venom for medicinal purposes dates back thousands of years. There are many reports in the literature on the pharmacological properties of bee venom and/or its main components, e.g., anti-arthritic, anti-inflammatory, anti-microbial or neuroprotective properties. In addition, both crude venom and melittin exhibit cytotoxic activity against a wide range of tumor cells, with significant anti-metastatic activity in pre-clinical studies. Due to the constantly increasing incidence of cancer, the development of new therapeutic strategies in oncology is a particular challenge for modern medicine. A review paper discusses the various properties of bee venom with an emphasis on its anticancer properties. For this purpose, the PubMed database was searched, and publications related to “bee”, “venom”, “cancer” from the last 10 years were selected.
Previous observations indicating a lower incidence of various types of cancer in beekeepers suggest that greater exposure to stings reduces the risk of cancer development. However, it is not known which of the active compounds of the bee venom (BV) may be responsible for the observed properties. The aim of this study is to evaluate the anti-glioblastoma effect of the main BV fractions. In addition, the effect of BV fractions on the activity of matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) was assessed. Commercially available BV was divided into three fractions containing one of the main BV components: apamin (fraction #1), phospholipase A2 (fraction #2), or melittin (fraction #3). The fraction containing apamin did not show cytotoxic activity up to a concentration of 100 µg/mL. The fraction containing phospholipase A2 partially reduced the cells’ viability at a concentration of 100 µg/mL. The greatest activity was demonstrated by the melittin-containing fraction which completely reduced the viability of glioma cells from a concentration of 2.5 μg/mL and inhibited the activity of the assessed metalloproteinases in a dose-dependent manner. After 72 h of incubation, the highest concentrations of TIMP-1 and TIMP-2 (approximately 150 ng/mL and 100 ng/mL, respectively) were observed in the LN229 line. In all tested lines, fraction #3, crude BV, and melittin reduced the secretion of both inhibitors into the medium in a dose-dependent manner. The melittin-containing fraction possessed anti-glioma properties in vitro, suggesting that melittin may be the main anticancer compound of BV.
Previous in vitro studies have shown the therapeutic potential of bee venom (BV) against different types of glioblastoma cells. Our aim was to evaluate the cytotoxic effect of BV on glioma in the zebrafish model. First, safe concentrations of BV and melittin were determined by determining the LD50 for each substance. Two human glioma cell lines, 8MGBA and LN-229, were used in this study. After staining the tested cells for visualization under UV light, they were then implanted into 2-day-old zebrafish embryos. Zebrafish were incubated for 3 days with crude BV and melittin at concentrations of 1.5 and 2.5 µg/mL vs. control group. Tumor growth was assessed with a stereo microscope. We found differential proliferative responses of two human glioma lines in a zebrafish model. The 8MGBA cell line, but not LN-229, showed proliferative potential when implanted into 2-day-old zebrafish embryos. This study showed a dose-dependent cytotoxic effect only for BV against 8MGBA cells. The observed cytotoxic effect is not dependent on the presence of the peptide melittin—the main BV component with the greatest cytotoxic potential. Simultaneously, a slight increase in LN-229 cell proliferation was observed after 3 days of incubation with melittin at a concentration of 2.5 µg/mL. This indicates that any consideration of bee venom as a therapeutic substance must take into account the type of glioblastoma.
https://lublin.tvp.pl/57079605/23-listopada-2021
https://radio.lublin.pl/2024/01/jad-pszczeli-w-walce-z-glejakiem-mozgu-badania-lubelskich-naukowcow/
Contact info: Department of Medical Chemistry, Medical University of Lublin, Chodzki 4a, 20-093 Lublin, Poland. email: beevenomproject@gmail.com