Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells

Aybüke Afra Babacan; Arif Ayar

doi:10.35206/jan.1808482

TR EN

Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells

Abstract

Breast cancer is one of the most common malignancies affecting women. Due to the side effects of conventional treatments, research into naturally occurring alternative therapeutic agents is critically important. This study aimed to comprehensively investigate, for the first time, the anticancer potential of lyophilized male bee larvae (apilarnil) and queen bee larvae, which are new generation apitherapy products, against breast cancer cells. The cytotoxic effects of apilarnil and queen bee larvae were evaluated in MCF-7 (human breast adenocarcinoma) and MCF-12a (healthy breast epithelium) cell lines using the MTT assay. Following the determination of IC₅₀ values, apoptotic cell death mechanisms were examined at the molecular level using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The study analyzed the mRNA expression levels of Bcl-2, Bax, Caspase-3, Caspase-8, and Caspase-9 genes, which play a critical role in apoptosis regulation. Both bee products exhibited potent cytotoxic activity against MCF-7 cancer cells (apilarnil IC₅₀: 7.7 µg/mL; queen bee larvae IC₅₀: 15.9 µg/mL). Molecular analyses revealed that these products increased the expression of proapoptotic genes (Bax, Caspase-3, Caspase-8, and Caspase-9) by 2-8-fold and significantly downregulated the anti-apoptotic Bcl-2 gene. A proliferative effect was observed in healthy MCF-12a cells at the same concentrations, indicating selective anticancer activity. This study demonstrates, for the first time, that apilarnil and queen bee larvae exhibit potent and selective cytotoxic effects against breast cancer cells. The findings indicate that these apitherapeutic products target cancer cells by activating apoptotic pathways. The results suggest that these natural products have potential as adjuvant therapies in future breast cancer treatments and provide a robust foundation for subsequent in vivo research in this field.

Keywords

Bee products, breast cancer, cell death, cytotoxicity, natural anticancer agents

Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells

Abstract

Breast cancer is one of the most common malignancies affecting women. Due to the side effects of conventional treatments, research into naturally occurring alternative therapeutic agents is critically important. This study aimed to comprehensively investigate, for the first time, the anticancer potential of lyophilized male bee larvae (apilarnil) and queen bee larvae, which are new generation apitherapy products, against breast cancer cells. The cytotoxic effects of apilarnil and queen bee larvae were evaluated in MCF-7 (human breast adenocarcinoma) and MCF-12a (healthy breast epithelium) cell lines using the MTT assay. Following the determination of IC₅₀ values, apoptotic cell death mechanisms were examined at the molecular level using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The study analyzed the mRNA expression levels of Bcl-2, Bax, Caspase-3, Caspase-8, and Caspase-9 genes, which play a critical role in apoptosis regulation. Both bee products exhibited potent cytotoxic activity against MCF-7 cancer cells (apilarnil IC₅₀: 7.7 µg/mL; queen bee larvae IC₅₀: 15.9 µg/mL). Molecular analyses revealed that these products increased the expression of proapoptotic genes (Bax, Caspase-3, Caspase-8, and Caspase-9) by 2-8-fold and significantly downregulated the anti-apoptotic Bcl-2 gene. A proliferative effect was observed in healthy MCF-12a cells at the same concentrations, indicating selective anticancer activity. This study demonstrates, for the first time, that apilarnil and queen bee larvae exhibit potent and selective cytotoxic effects against breast cancer cells. The findings indicate that these apitherapeutic products target cancer cells by activating apoptotic pathways. The results suggest that these natural products have potential as adjuvant therapies in future breast cancer treatments and provide a robust foundation for subsequent in vivo research in this field.

Keywords

Bee products, breast cancer, cell death, cytotoxicity, natural anticancer agents

Ethical Statement

The authors of this article declare that the materials and methods used in this study do not require ethical committee approval and/or legal-specific permission.

References

Abd El‐Wahed, A. A., Khalifa, S. A., Aldahmash, B., Zhang, H., Du, M., Zhao, C., . . . & El‐Seedi, H. R. (2024). Exploring the chemical constituents and nutritive potential of bee drone (Apilarnil): emphasis on antioxidant properties. Chemistry and Biodiversity, 21(5), e202400085. https://doi.org/10.1002/cbdv.202400085
American Cancer Society. (2018). What causes cancer? https://www.cancer.org/cancer/cancercauses.html
Aoşan, C. (2016). Apitherapy in the daily practice — Clinical applications [Conference presentation]. Apimedica and Apiquality Forum, Rome, Italy.
Arnold, M., Morgan, E., Rumgay, H., Mafra, A., Singh, D., Laversanne, M., . . . & Soerjomataram, I. (2022). Current and future burden of breast cancer: Global statistics for 2020 and 2040. The Breast, 66, 15–23. https://doi.org/10.1016/j.breast.2022.08.010
Babacan, A. A., & Ayar, A. (2024). Effects of apilarnil and queen bee larvae on larval mortality and longevity in Drosophila melanogaster. Uludağ Arıcılık Dergisi, 24(2), 223–234. https://doi.org/10.31467/uluaricilik.1494204
Badisa, R. B., Darling-Reed, S. F., Joseph, P., Cooperwood, J. S., Latinwo, L. M., & Goodman, C. B. (2009). Selective cytotoxic activities of two novel synthetic drugs on human breast carcinoma MCF-7 cells. Anticancer Research, 29(8), 2993–2996.
Balkanska, R., Karadjova, I., & Ignatova, M. (2014). Comparative analyses of chemical composition of royal jelly and drone brood. Bulgarian Chemical Communications, 46(2), 412–416.
Bărnuţiu, L. I., Mărghitaş, L. A., Dezmirean, D., Bobiş, O., Mihai, C., & Pavel, C. (2013). Physico-chemical composition of apilarnil (bee drone larvae). Lucrări Ştiinţifice—Seria Zootehnie, 59, 199–202.
Borum, A. E. (2024). Apitherapy and applications in veterinary medicine. Uludağ Arıcılık Dergisi, 24(2), 442–458. https://doi.org/10.31467/uluaricilik.1537629
Cacan, E., Spring, A. M., Kumari, A., Greer, S. F., & Garnett-Benson, C. (2015). Combination treatment with sublethal ionizing radiation and the proteasome inhibitor, bortezomib, enhances death-receptor mediated apoptosis and anti-tumor immune attack. International Journal of Molecular Sciences, 16(12), 30405–30421. https://doi.org/10.3390/ijms161226238

Calabrese, E. J. (2008). Hormesis: Why it is important to toxicology and toxicologists. Environmental Toxicology and Chemistry, 27(7), 1451–1474. https://doi.org/10.1897/07-541.1
Calabrese, E. J., & Baldwin, L. A. (2003). Hormesis: The dose-response revolution. Annual Review of Pharmacology and Toxicology, 43(1), 175–197. https://doi.org/10.1146/annurev.pharmtox.43.100901.140223
Çelik, K., & Aşgun, H. F. (2020). Apitherapy: Health and healing from the bees. Tudás Alapítvány.
Comşa, Ş., Cimpean, A. M., & Raica, M. (2015). The story of MCF-7 breast cancer cell line: 40 years of experience in research. Anticancer Research, 35(6), 3147–3154.
Doğanyiğit, Z., Okan, A., Kaymak, E., Pandır, D., & Silici, S. (2020). Investigation of protective effects of apilarnil against lipopolysaccharide induced liver injury in rats via TLR 4/HMGB-1/NF-κB pathway. Biomedicine and Pharmacotherapy, 125, 109967. https://doi.org/10.1016/j.biopha.2020.109967
Duffy, C., Sorolla, A., Wang, E., Golden, E., Woodward, E., Davern, K., . . . & Blancafort, P. (2020). Honeybee venom and melittin suppress growth factor receptor activation in HER2-enriched and triple-negative breast cancer. NPJ Precision Oncology, 4(1), 24. https://doi.org/10.1038/s41698-020-00129-0
El-Garawani, I., El-Seedi, H., Khalifa, S., El Azab, I. H., Abouhendia, M., & Mahmoud, S. (2020). Enhanced antioxidant and cytotoxic potentials of lipopolysaccharides-injected Musca domestica larvae. Pharmaceutics, 12(11), 1111. https://doi.org/10.3390/pharmaceutics12111111
Erdem, B., & Özkök, A. (2018). Can food supplement produced from apilarnil be an alternative to testosterone replacement therapy? Hacettepe Journal of Biology and Chemistry, 45(4), 635–638.
Fratini, F., Cilia, G., Mancini, S., & Felicioli, A. (2016). Royal jelly: An ancient remedy with remarkable antibacterial properties. Microbiological Research, 192, 130–141. https://doi.org/10.1016/j.micres.2016.06.007
Freshney, R. I. (2015). Culture of animal cells: A manual of basic technique and specialized applications. John Wiley and Sons.
Ganot, N., Meker, S., Reytman, L., Tzubery, A., & Tshuva, E. Y. (2013). Anticancer metal complexes: Synthesis and cytotoxicity evaluation by the MTT assay. Journal of Visualized Experiments, 81, 50767. https://doi.org/10.3791/50767
Gavrila, M. A. (2010). The concepts of normality, health-abnormal, disease. Agora, 4(15), 29–36.
Gautam, A., Gyawali, I., Poudel, S., Devkota, S., Acharya, R., Kandel, M., & Subedi, D. (2025). Insects as food and feed source: A comprehensive review on nutritional value, food safety concern, environmental benefits, economic potential, technological innovations, challenges, and future prospects. Food Frontiers, 6(6), 2591–2646. https://doi.org/10.1002/fft2.70092
Giaquinto, A. N., Sung, H., Newman, L. A., Freedman, R. A., Smith, R. A., Star, J., . . . & Siegel, R. L. (2024). Breast cancer statistics 2024. CA: A Cancer Journal for Clinicians, 74(6), 477–495. https://doi.org/10.3322/caac.21863
Gökkaya, İ., Renda, G., Ayhan, F., & Yazici, N. (2025). Awareness of apitherapy: A study of healthcare professionals in Türkiye. Journal of Pharmacopuncture, 28(1), 35. https://doi.org/10.3831/KPI.2025.28.1.35
Hamamci, M., Doganyigit, Z., Silici, S., Okan, A., Kaymak, E., Yilmaz, S., . . . & Inan, L. E. (2020). Apilarnil: A novel neuroprotective candidate. Acta Neurologica Taiwanica, 29(2), 33–45. Isidorov, V. A., Bakier, S., & Stocki, M. (2016). GC-MS investigation of the chemical composition of honeybee drone and queen larvae homogenate. Journal of Apicultural Science, 60(2), 4–16. https://doi.org/10.1515/JAS-2016-0011
Kalkan, K. T., Yildiz, H. T., Yalçin, B., Mat, Ö. C., Önder, G. Ö., & Yay, A. (2025). Therapeutic effects of apilarnil, a bee product, on apoptosis and autophagy in cisplatin-induced rat ovarian toxicity. Food and Chemical Toxicology, 200, 115594. https://doi.org/10.1016/j.fct.2025.115594
Kanyilmaz, G., Akmansu, M., Bora, H., & Yirmibeşoğlu Erkal, E. D. A. (2022). The use of complementary and alternative medicine among cancer patients treated with radiotherapy. Turkish Journal of Oncology, 37(1). https://doi.org/10.5505/tjo.2021.3357
Kerr, J. F., Wyllie, A. H., & Currie, A. R. (1972). Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics. British Journal of Cancer, 26(4), 239–257. https://doi.org/10.1038/bjc.1972.33
Koc, K., Erol, H. S., Colak, S., Cerig, S., Yildirim, S., & Geyikoglu, F. (2019). The protective effect of propolis on rat ovary against ischemia-reperfusion injury: Immunohistochemical, biochemical and histopathological evaluations. Biomedicine and Pharmacotherapy, 111, 631–637. https://doi.org/10.1016/j.biopha.2018.12.113
Kogalniceanu, S., Lancrajan, I., & Ardelean, G. (2010). Changes of the glucidic metabolism determined by the physical effort of the treatment with the Aslavital and Apilarnil. Journal of the Medical Association of Arad, 3, 33–41.
Kumariya, S., Ubba, V., Jha, R. K., & Gayen, J. R. (2021). Autophagy in ovary and polycystic ovary syndrome: Role, dispute and future perspective. Autophagy, 17(10), 2706–2733. https://doi.org/10.1080/15548627.2021.1938914
Mărgăoan, R., Mărghitaş, L. A., Dezmirean, D. S., Bobiş, O., Bonta, V., Cătană, C., . . . & Margin, M. G. (2017). Comparative study on quality parameters of royal jelly, apilarnil and queen bee larvae triturate. Bulletin of the University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca. Animal Science and Biotechnologies, 74(1). https://doi.org/10.15835/buasvmcn-asb:12622
Nadendla, E. K., Tweedell, R. E., Kasof, G., & Kanneganti, T. D. (2025). Caspases: Structural and molecular mechanisms and functions in cell death, innate immunity, and disease. Cell Discovery, 11(1), 42. https://doi.org/10.1038/s41421-025-00791-3
Omar, W. A. W., Azhar, N. A., Fadzilah, N. H., & Kamal, N. N. S. N. M. (2016). Bee pollen extract of Malaysian stingless bee enhances the effect of cisplatin on breast cancer cell lines. Asian Pacific Journal of Tropical Biomedicine, 6(3), 265–269. https://doi.org/10.1016/j.apjtb.2015.12.011
Rumpold, B. A., & Schlüter, O. K. (2013). Nutritional composition and safety aspects of edible insects. Molecular Nutrition and Food Research, 57(5), 802–823. https://doi.org/10.1002/mnfr.201200735
Rutka, I., Galoburda, R., Galins, J., & Galins, A. (2021). Bee drone brood homogenate chemical composition, stabilization and application: A review. Research for Rural Development, 36, 96–103. https://doi.org/10.22616/rrd.27.2021.014
Sanlier, N., Kaya, E. Y., & Yucel, I. I. (2026). Health effects of bee products: A comprehensive review. Food Science and Nutrition, 14(2), e71165. https://doi.org/10.1002/fsn3.71165
Sawczuk, R., Karpinska, J., & Miltyk, W. (2019). What do we need to know about drone brood homogenate and what is known. Journal of Ethnopharmacology, 245, 111581. https://doi.org/10.1016/j.jep.2018.10.042
Schmittgen, T. D., & Livak, K. J. (2008). Analyzing real-time PCR data by the comparative C(T) method. Nature Protocols, 3(6), 1101–1108. https://doi.org/10.1038/nprot.2008.73
Sidor, E., & Dżugan, M. (2020). Drone brood homogenate as natural remedy for treating health care problem: A scientific and practical approach. Molecules, 25(23), 5699. https://doi.org/10.3390/molecules25235699
Silici, S. (2023). Drone larvae homogenate (Apilarnil) as natural remedy: Scientific review. Journal of Agricultural Sciences, 29(4), 947–959. https://doi.org/10.15832/ankutbd.1293015
Strant, M., Varadi, A., & Aoşan, C. (2016). The apilarnil and queen larvae—Studies, utilization, doses, clinical cases. Cluj Napoca.
Sulaiman, C., George, B. P., Balachandran, I., & Abrahamse, H. (2025). Cancer and traditional medicine: An integrative approach. Pharmaceuticals, 18(5), 644. https://doi.org/10.3390/ph18050644
Topal, E., Strant, M., Yücel, B., Kösoğlu, M., Margaoan, R., & Dayıoğlu, M. (2018). Ana ve erkek arı larvalarının biyokimyasal özellikleri ve apiterapötik kullanımı. Journal of Animal Production, 59(2), 77–82. https://doi.org/10.29185/hayuretim.455478
Van Huis, A., Dicke, M., & Van Loon, J. J. (2015). Insects to feed the world. Journal of Insects as Food and Feed, 1(1), 3–6. https://doi.org/10.3920/JIFF2015.x002
Yılmaz, E. A., & Babacan, A. A. (2024). Apilarnil ve kraliçe arı larvası liyofilizatlarının genotoksik ve antigenotoksik etkilerinin mikronükleus testi ile belirlenmesi. Gazi Üniversitesi Fen Fakültesi Dergisi, 5(1), 11–20.
Yuan, L., Cai, Y., Zhang, L., Liu, S., Li, P., & Li, X. (2022). Promoting apoptosis, a promising way to treat breast cancer with natural products: A comprehensive review. Frontiers in Pharmacology, 12, 801662. https://doi.org/10.3389/fphar.2021.801662
Zedan, A. M., Sakran, M. I., Bahattab, O., Hawsawi, Y. M., Al-Amer, O., Oyouni, A. A., . . . & El-Magd, M. A. (2021). Oriental Hornet (Vespa orientalis) larval extracts induce antiproliferative, antioxidant, anti-inflammatory, and anti-migratory effects on MCF7 cells. Molecules, 26(11), 3303. https://doi.org/10.3390/molecules26113303

Details

Primary Language

English

Subjects

Metabolomic Chemistry

Journal Section

Research Article

Authors

Aybüke Afra Babacan
0000-0003-4333-4732
Türkiye

Arif Ayar ^*
0000-0003-0473-4653
Türkiye

Publication Date

April 27, 2026

Submission Date

October 22, 2025

Acceptance Date

December 27, 2025

Published in Issue

Year 2026 Volume: 9 Number: 1

DOI

https://doi.org/10.35206/jan.1808482

IZ

https://izlik.org/JA36JJ66HP

APA

Babacan, A. A., & Ayar, A. (2026). Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells. Journal of Apitherapy and Nature, 9(1), 22-36. https://doi.org/10.35206/jan.1808482

AMA

1.Babacan AA, Ayar A. Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells. J.Apit.Nat. 2026;9(1):22-36. doi:10.35206/jan.1808482

Chicago

Babacan, Aybüke Afra, and Arif Ayar. 2026. “Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells”. Journal of Apitherapy and Nature 9 (1): 22-36. https://doi.org/10.35206/jan.1808482.

EndNote

Babacan AA, Ayar A (April 1, 2026) Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells. Journal of Apitherapy and Nature 9 1 22–36.

IEEE

[1]A. A. Babacan and A. Ayar, “Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells”, J.Apit.Nat., vol. 9, no. 1, pp. 22–36, Apr. 2026, doi: 10.35206/jan.1808482.

ISNAD

Babacan, Aybüke Afra - Ayar, Arif. “Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells”. Journal of Apitherapy and Nature 9/1 (April 1, 2026): 22-36. https://doi.org/10.35206/jan.1808482.

JAMA

1.Babacan AA, Ayar A. Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells. J.Apit.Nat. 2026;9:22–36.

MLA

Babacan, Aybüke Afra, and Arif Ayar. “Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells”. Journal of Apitherapy and Nature, vol. 9, no. 1, Apr. 2026, pp. 22-36, doi:10.35206/jan.1808482.

Vancouver

1.Aybüke Afra Babacan, Arif Ayar. Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells. J.Apit.Nat. 2026 Apr. 1;9(1):22-36. doi:10.35206/jan.1808482

e-ISSN: 2667-4734 Publication Model: Continuous Publication Citation Indexes: TR Dizin

Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells

Abstract

Keywords

Molecular Investigation of the Cytotoxic and Apoptotic Effects of Apilarnil and Queen Bee Larvae in MCF-7 Breast Cancer Cells

Abstract

Keywords

Ethical Statement

References

Details

Primary Language

Subjects

Journal Section

Authors

Publication Date

Submission Date

Acceptance Date

Published in Issue

DOI

IZ

Cite