Investigation of apoptotic efficacy of propolis in MCF-7 cell line

Year 2024, Volume: 15 Issue: 52, 80 - 85, 02.09.2024

Gülay Gülbol Duran

https://doi.org/10.17944/interdiscip.1466355

Abstract

Objective: Propolis, also known as bee glue, is a resinous compound collected by honey bees from various plants and processed by their saliva enzymes. Propolis and its components have been studied for their cytotoxic effects on cell lines in vitro, and recent studies have shown that they also have an antitumor effect in vivo. This study aimed to investigate the in-vitro apoptotic effects of propolis on the human breast cancer cell line (MCF-7).
Method: The MTT test was used to determine the effect of propolis on cell viability and the doses to be administered. The GraphPad Prism Version 6.01 program was used to analyze the MTT results, while the qRT-PCR method was used to determine the expression levels of Caspase-8, Caspase-9, and Bcl-2 genes. The RT2 profiler PCR Assay Data Analysis version 3.5 was used to analyze gene expression data.
Results: This study it was found that doses of 3.9 and 7.8 µg/ml of propolis showed no cytotoxic effect, while doses of 15.625 µg/ml and above had a cytotoxic effect. There was no change in the expression levels of genes at concentrations of 3.9 µg/ml and 7.8 µg/ml of propolis. However, at 15.625 µg/ml of propolis, Caspase-9 gene expression increased 11.89-fold (p=0.033). Although there was no significant difference in Caspase-8 gene expression in the extrinsic pathway of apoptosis (p=0.437), a 0.04-fold decrease in anti-apoptotic Bcl-2 gene expression was observed (p=0.000098).
Conclusion: In conclusion, propolis showed a dose-dependent cytotoxic effect on the MCF-7 cell line, induced apoptosis, and did so via the intrinsic pathway of apoptosis. The study suggests that propolis has high potential as an anticancer agent since its apoptotic effects have been demonstrated in the MCF-7 cell line.

Keywords

Propolis, MCF-7, Apoptosis, Caspase-9, Bcl-2

References

• Zaccaria V, Curti V, Di Lorenzo A, Baldi A, Maccario C, Sommatis S, et al. Effect of Green and Brown Propolis Extracts on the Expression Levels of microRNAs, mRNAs and Proteins, Related to Oxidative Stress and Inflammation. Nutrients. 2017;9(10):1090. doi: 10.3390/nu9101090.
• Vukovic NL, Obradovic AD, Vukic MD, Jovanovic D, Djurdjevic PM. Cytotoxic, proapoptotic and antioxidative potential of flavonoids isolated from propolis against colon (HCT-116) and breast (MDA-MB-231) cancer cell lines. Food Res Int. 2018;106:71-80. doi: 10.1016/j.foodres.2017.12.056.
• Tsuda T, Kumazawa S. Propolis: Chemical Constituents, Plant Origin, and Possible Role in the Prevention and Treatment of Obesity and Diabetes. J Agric Food Chem. 2021;69(51):15484-15494. doi: 10.1021/acs.jafc.1c06194.
• Freitas AS, Cunha A, Oliveira R, Almeida-Aguiar C. Propolis antibacterial and antioxidant synergisms with gentamicin and honey. J Appl Microbiol. 2022;132(4):2733-2745. doi: 10.1111/jam.15440.
• Oliveira JMDS, Cavalcanti TFS, Leite IF, Dos Santos DMRC, Porto ICCM, de Aquino FLT et al. Propolis in Oral Healthcare: Antibacterial Activity of a Composite Resin Enriched With Brazilian Red Propolis. Front Pharmacol. 2021;12:787633. doi: 10.3389/fphar.2021.787633.
• Wang F, Liu H, Li J, Zhang W, Jiang B, Xuan H. Australian propolis ethanol extract exerts antibacterial activity against methicillin-resistant Staphylococcus aureus by mechanisms of disrupting cell structure, reversing resistance, and resisting biofilm. Braz J Microbiol. 2021;52(4):1651-1664. doi: 10.1007/s42770-021-00547-7.
• Xu W, Lu H, Yuan Y, Deng Z, Zheng L, Li H. The Antioxidant and Anti-Inflammatory Effects of Flavonoids from Propolis via Nrf2 and NF-κB Pathways. Foods. 2022;11(16):2439. doi: 10.3390/foods11162439.
• Papp Z, Bouchelaghem S, Szekeres A, Meszéna R, Gyöngyi Z, Papp G. The Scent of Antifungal Propolis. Sensors (Basel). 2021;21(7):2334. doi: 10.3390/s21072334.
• Santos LA, Rosalen PL, Dias NA, Grisolia JC, Nascimento Gomes BJ, Blosfeld-Lopes L et al. Brazilian Red Propolis shows antifungal and immunomodulatory activities against Paracoccidioides brasiliensis. J Ethnopharmacol. 2021 Sep 15;277:114181. doi: 10.1016/j.jep.2021.114181.
• Yildirim A, Duran GG, Duran N, Jenedi K, Bolgul BS, Miraloglu M et al. Antiviral Activity of Hatay Propolis Against Replication of Herpes Simplex Virus Type 1 and Type 2. Med Sci Monit. 2016;22:422-30. doi: 10.12659/msm.897282.
• Demir S, Atayoglu AT, Galeotti F, Garzarella EU, Zaccaria V, Volpi N et al. Antiviral activity of different extracts of standardized propolis preparations against HSV. Antivir Ther. 2020;25(7):353-363. doi: 10.3851/IMP3383.
• Silva-Beltrán NP, Balderrama-Carmona AP, Umsza-Guez MA, Souza Machado BA. Antiviral effects of Brazilian green and red propolis extracts on Enterovirus surrogates. Environ Sci Pollut Res Int. 2020;27(23):28510-28517. doi: 10.1007/s11356-019-07458-z.
• Salem MM, Donia T, Abu-Khudir R, Ramadan H, Ali EMM, Mohamed TM. Propolis Potentiates Methotrexate Anticancer Mechanism and Reduces its Toxic Effects. Nutr Cancer. 2020;72(3):460-480. doi: 10.1080/01635581.2019.1640884.
• Alanazi S, Alenzi N, Alenazi F, Tabassum H, Watson D. Chemical characterization of Saudi propolis and its antiparasitic and anticancer properties. Sci Rep. 2021;11(1):5390. doi: 10.1038/s41598-021-84717-5.
• Rouibah H, Kebsa W, Lahouel M, Zihlif M, Ahram M, Aburmaileh B et al. Algerian propolis: between protection of normal cells and potentialisation of the anticancer effects of doxorubicin against breast cancer cells via P-glycoprotein inhibition and cell cycle arrest in the S phase. J Physiol Pharmacol. 2021;72(2). doi: 10.26402/jpp.2021.2.09.
• Ishida Y, Gao R, Shah N, Bhargava P, Furune T, Kaul SC et al. Anticancer Activity in Honeybee Propolis: Functional Insights to the Role of Caffeic Acid Phenethyl Ester and Its Complex With γ-Cyclodextrin. Integr Cancer Ther. 2018;17(3):867-873. doi: 10.1177/1534735417753545.
• Memmedov H, Oktay LM, Durmaz B, Günel NS, Yıldırım HK, Sözmen EY. Propolis prevents inhibition of apoptosis by potassium bromate in CCD 841 human colon cell. Cell Biochem Funct. 2020;38(4):510-519. doi: 10.1002/cbf.3475.
• Salehi A, Hosseini SM, Kazemi S. Antioxidant and Anticarcinogenic Potentials of Propolis for Dimethylhydrazine-Induced Colorectal Cancer in Wistar Rats. Biomed Res Int. 2022;2022:8497562. doi: 10.1155/2022/8497562.
• Moskwa J, Naliwajko SK, Markiewicz-Żukowska R, Gromkowska-Kępka KJ, Soroczyńska J, Puścion-Jakubik A et al. Polish and New Zealand Propolis as Sources of Antioxidant Compounds Inhibit Glioblastoma (T98G, LN-18) Cell Lines and Astrocytoma Cells Derived from Patient. Antioxidants (Basel). 2022;11(7):1305. doi: 10.3390/antiox11071305.
• Jiang XS, Xie HQ, Li CG, You MM, Zheng YF, Li GQ et al. Chinese Propolis Inhibits the Proliferation of Human Gastric Cancer Cells by Inducing Apoptosis and Cell Cycle Arrest. Evid Based Complement Alternat Med. 2020;2020:2743058. doi: 10.1155/2020/2743058.
• Coskun ZM, Ersoz M, Gecili M, Ozden A, Acar A. Cytotoxic and apoptotic effects of ethanolic propolis extract on C6 glioma cells. Environ Toxicol. 2020;35(7):768-773. doi: 10.1002/tox.22911.
• Sahin U, Türeci Ö. Personalized vaccines for cancer immunotherapy. Science. 2018;359(6382):1355-1360. doi: 10.1126/science.aar7112.
• Judasz E, Lisiak N, Kopczyński P, Taube M, Rubiś B. The Role of Telomerase in Breast Cancer's Response to Therapy. Int J Mol Sci. 2022;23(21):12844. doi: 10.3390/ijms232112844.
• Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71(3):209-249. doi: 10.3322/caac.21660.
• Morana O, Wood W, Gregory CD. The Apoptosis Paradox in Cancer. Int J Mol Sci. 2022;23(3):1328. doi: 10.3390/ijms23031328.
• Kashyap D, Garg VK, Goel N. Intrinsic and extrinsic pathways of apoptosis: Role in cancer development and prognosis. Adv Protein Chem Struct Biol. 2021;125:73-120. doi: 10.1016/bs.apcsb.2021.01.003.
• de Mendonça IC, Porto IC, do Nascimento TG, de Souza NS, Oliveira JM, Arruda RE et al. Brazilian red propolis: phytochemical screening, antioxidant activity and effect against cancer cells. BMC Complement Altern Med. 2015;15:357. doi: 10.1186/s12906-015-0888-9.
• Masadah R, Ikram D, Rauf S. Effects of propolis and its bioactive components on breast cancer cell pathways and the molecular mechanisms involved. Breast Dis. 2021;40(S1):S15-S25. doi: 10.3233/BD-219003.
• [Internet]:https://hsgm.saglik.gov.tr/depo/birimler/kanserdb/Dokumanlar/Istatistikler/Kanser_Rapor_2018.pdf. (cited:16.03.2024).
• Bulut S, Yaylım İ. SLAMF7:A Novel Cancer-Related Target Molecule. Journal of Medical Sciences.2024,5(1)1-7. https://doi.org/10.46629/JMS.2024.138
• Meghalatha TS, Muninathan N. Antitumor activity of withaferin-A and propolis in benz (a) pyrene-induced breast cancer. Bioinformation. 2022;18(9):841-844. doi: 10.6026/97320630018841.
• Tsuda T, Kumazawa S. Propolis: Chemical Constituents, Plant Origin, and Possible Role in the Prevention and Treatment of Obesity and Diabetes. J Agric Food Chem. 2021;69(51):15484-15494. doi: 10.1021/acs.jafc.1c06194.
• Amalia E, Diantini A, Subarnas A. Water-soluble propolis and bee pollen of Trigona spp. from South Sulawesi Indonesia induce apoptosis in the human breast cancer MCF-7 cell line. Oncol Lett. 2020;20(5):274. doi: 10.3892/ol.2020.12137. Epub 2020 Sep 21.
• Xuan H, Li Z, Yan H, Sang Q, Wang K, He Q et al. Antitumor Activity of Chinese Propolis in Human Breast Cancer MCF-7 and MDA-MB-231 Cells. Evid Based Complement Alternat Med. 2014;2014:280120. doi: 10.1155/2014/280120.
• Seyhan MF, Yılmaz E, Timirci-Kahraman Ö, Saygılı N, Kısakesen Hİ, Gazioğlu S et al. Different propolis samples, phenolic content, and breast cancer cell lines: Variable cytotoxicity ranging from ineffective to potent. IUBMB Life. 2019;71(5):619-631. doi: 10.1002/iub.1996.
• Vatansever SH, Sorkun K, Ismet Deliloğlu Gurhan S, Ozdal-Kurt F, Turkoz E, Gencay O, Salih B. Propolis from Turkey induces apoptosis through activating caspases in human breast carcinoma cell lines. Acta Histochem. 2010;112(6):546-56. doi: 10.1016/j.acthis.2009.06.001.
• Misir S, Aliyazicioglu Y, Demir S, Turan I, Hepokur C. Effect of Turkish Propolis on miRNA Expression, Cell Cycle, and Apoptosis in Human Breast Cancer (MCF-7) Cells. Nutr Cancer. 2020;72(1):133-145. doi: 10.1080/01635581.2019.1616100.
• Niyomtham N, Koontongkaew S, Yingyongnarongkul BE, Utispan K. Apis melliferapropolis enhances apoptosis and invasion inhibition in head and neck cancer cells. PeerJ. 2021;9:e12139. doi: 10.7717/peerj.12139.