Propolis: Intrinsic Pathway-Induced Apoptosis, G1 Cell Cycle Arrest, Reduced Chemotherapeutic Resistance in Adenocarcinoma, and Healthy Cell Preservation

Year 2023, , 194 - 204, 28.12.2023

Nursel İşçi , Sibel Doğan , Sahika Yardım , Seyhun Solakoğlu , İlhan Tahralı , Günnur Deniz , Gülnaz Nural Bekiroğlu , Elif Kervancıoğlu Demirci

https://doi.org/10.26650/experimed.1343683

Abstract

Objective: This study investigated the potential synergistic effects of propolis, an antitumor and antioxidant natural product, and carboplatin, a frequently used chemotherapeutic agent for endometrial adenocarcinoma, one of the most common gynaecological cancers treatment.
Materials and Methods: Ishikawa endometrial adenocarcinoma and healthy fibroblast (3T3) cell lines were treated with 0.5 µL of carboplatin and 5 µL of propolis. Cell count, viability, migration, ultrastructure, apoptosis, and cell cycle changes were assessed using cytological and immunocytochemical methods, flow cytometry, and transmission electron microscopy (TEM).
Results: Propolis and carboplatin exhibited cytotoxic effects on Ishikawa cells. The combination of the two agents further reduced cell viability and migration. Propolis induced apoptosis through the intrinsic pathway and arrested the cell cycle in the G1 phase. TEM analysis revealed apoptosis in Ishikawa cells treated with propolis or carboplatin, while the carboplatin+propolis combination resulted in severe cell budding, apoptosis, and vacuolization. Migrasome-like structures were only observed in the Ishikawa carboplatin group. Minimal effects were observed on 3T3 cells.
Conclusion: Propolis demonstrated cytotoxic, anti-proliferative, and proapoptotic effects on tumor cells without harming healthy cells. Its ability to prevent migrasome formation suggests it may reduce chemotherapeutic resistance. Therefore, propolis shows promise as a potential enhancer of anticancer treatments.

Keywords

3T3, apoptosis, electron microscopy, endometrium, Ishikawa, migrasome

Supporting Institution

İstanbul Üniversitesi Bilimsel Araştırma Projeleri Birimi

Project Number

37221

Thanks

This study was supported by Istanbul University Scientific Research Projects Unit (Project No: 37221)

References

• 1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin 2011; 61(2): 69-90. google scholar
• 2. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin 2016; 66(1): 7-30. google scholar
• 3. Crissman JD, Azoury RS, Barnes AE, Schellhas HF. Endometrial carcinoma in women 40 years of age or younger. Obstet Gynecol 1981; 57(6): 699-704. google scholar
• 4. Nishida M. The Ishikawa cells from birth to the present. Hum Cell 2002; 15(3): 104-17. google scholar
• 5. Woodford-Thomas TA, Rhodes JD, Dixon JE. Expression of a protein tyrosine phosphatase in normal and v-src-transformed mouse 3T3 fibroblasts. J Cell Biol 1992; 117(2): 401-14. google scholar
• 6. Zagouri F, Bozas G, Kafantari E, Tsiatas M, Nikitas N, Dimopoulos MA, et al. Endometrial cancer: what is new in adjuvant and molecularly targeted therapy? Obstet Gynecol Int 2010; 2010: 749579. google scholar
• 7. Kumar A, Sirohi VK, Anum F, Singh PK, Gupta K, Gupta D, et al. Enhanced apoptosis, survivin down-regulation and assisted immunochemotherapy by curcumin-loaded amphiphilic mixed micelles for subjugating endometrial cancer. Nanomedicine 2017; 13(6): 1953-63. google scholar
• 8. SGO Clinical Practice Endometrial Cancer Working Group; Burke WM, Orr J, Leitao M, Salom E, Gehrig P, et al. Endometrial cancer: a review and current management strategies: part II. Gynecol Oncol 2014; 134(2): 393-402. google scholar
• 9. Jia N, Che X, Jiang Y, Zhu M, Yang T, Feng W. Synergistic effects of a combined treatment of PI3K/mTOR dual inhibitor LY3023414 and carboplatin on human endometrial carcinoma. Gynecol Oncol 2021; 162(3): 788-96. google scholar
• 10. Castaldo S, Capasso F. Propolis, an old remedy used in modern medicine. Fitoterapia 2002; 73 Suppl 1:S1-6. google scholar
• 11. Burdock GA. Review of the biological properties and toxicity of bee propolis (propolis). Food Chem Toxicol 1998; 36(4): 347-63. google scholar
• 12. Viuda Martos M, Ruiz Navajas Y, Fernandez Lopez J, Perez Âlvarez JA. Functional properties of honey, propolis, and royal jelly. J Food Sci 2008; 73(9): R117-24. google scholar
• 13. Turan I, Demir S, Misir S, Kilinc K, Mentese A, Aliyazicioglu Y, et al. Cytotoxic effect of Turkish propolis on liver, colon, breast, cervix and prostate cancer cell lines. Trop J Pharm Res 2015; 14(5): 77782. google scholar
• 14. Demir S, Aliyazicioglu Y, Turan I, Misir S, Mentese A, Yaman SO, et al. Antiproliferative and proapoptotic activity of Turkish propolis on human lung cancer cell line. Nutr Cancer 2016; 68(1): 165-72. google scholar
• 15. Kubina R, Kabala-Dzik A, Dziedzic A, Bielec B, Wojtyczka RD, Buldak RJ, et al. The ethanol extract of Polish propolis exhibits anti-proliferative and/or pro-apoptotic effect on HCT 116 colon cancer and Me45 malignant melanoma cells in vitro conditions. Adv Clin Exp Med 2015; 24(2): 203-12. google scholar
• 16. Rantanen V, Grenman S, Kulmala J, Grenman R. Comparative evaluation of cisplatin and carboplatin sensitivity in endometrial adenocarcinoma cell lines. Br J Cancer 1994; 69(3): 482-6. google scholar
• 17. Kuittinen T, Rovio P, Staff S, Luukkaala T, Kallioniemi A, Grenman S, et al. Paclitaxel, carboplatin and 1,25-D3 inhibit proliferation of endometrial cancer cells in vitro. Anticancer Res 2017; 37(12): 6575-81. google scholar
• 18. Lee CS, Kim YJ, Jang ER, Myung SC, Kim W. Akt inhibitor enhances apoptotic effect of carboplatin on human epithelial ovarian carcinoma cell lines. Eur J Pharmacol 2010; 632(1-3): 7-13. google scholar
• 19. Singh S, Upadhyay AK, Ajay AK, Bhat MK. p53 regulates ERK activation in carboplatin induced apoptosis in cervical carcinoma: a novel target of p53 in apoptosis. FEBS Lett 2007; 581(2): 289-95. google scholar
• 20. He PJ, Ge RF, Mao WJ, Chung PS, Ahn JC, Wu HT. Oxidative stress induced by carboplatin promotes apoptosis and inhibits migration of HN-3 cells. Oncol Lett 2018; 16(6): 7131-8. google scholar
• 21. Coleman SC, Stewart ZA, Day TA, Netterville JL, Burkey BB, Pietenpol JA. Analysis of cell-cycle checkpoint pathways in head and neck cancer cell lines: implications for therapeutic strategies. Arch Otolaryngol Head Neck Surg 2002; 128(2): 167-76. google scholar
• 22. Kelland L The resurgence of platinum-based cancer chemotherapy Nat Rev Cancer 2007; 7(8): 573-84 google scholar
• 23. Sousa GFd, Wlodarczyk SR, Monteiro G Carboplatin: molecular mechanisms of action associated with chemoresistance Brazilian J Pharma Sci 2014; 50: 693-701 google scholar
• 24. Eguchi T, Taha EA, Calderwood SK, Ono K A novel model of cancer drug resistance: oncosomal release of cytotoxic and antibody-based drugs Biology (Basel) 2020; 9(3): 47 google scholar
• 25. Eguchi T, Ono K, Calderwood S, Okamoto K Exosome release of drugs: coupling with epithelial-mesenchymal transition Preprints 2019; 2019120386 doi:10 20944/preprints201912 0386 v1 google scholar
• 26. Zhao X, Wu X, Qian M, Song Y, Wu D, Zhang W Knockdown of TGF-beta1 expression in human umbilical cord mesenchymal stem cells reverts their exosome-mediated EMT promoting effect on lung cancer cells Cancer Lett 2018; 428: 34-44 google scholar
• 27. Orsolic N, Knezevic AH, Sver L, Terzic S, Basic I Immunomodulatory and antimetastatic action of propolis and related polyphenolic compounds J Ethnopharmacol 2004; 94(2-3): 307-15 google scholar
• 28 . Lamoral-Theys D, Pottier L, DufrasneF, NeveJ, DuboisJ, Kornienko A, et al. Natural polyphenols that display anticancer properties through inhibition of kinase activity. Curr Med Chem 2010; 17(9):812- 25. google scholar
• 29. 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. google scholar
• 30. Desamero MJ, Kakuta S, Tang Y, Chambers JK, Uchida K, Estacio MA, et al. Tumor-suppressing potential of stingless bee propolis in in vitro and in vivo models of differentiated-type gastric adenocarcinoma. Sci Rep 2019; 9(1): 19635. google scholar
• 31. 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): 133-45. google scholar
• 32. Szliszka E, Czuba ZP, Domino M, Mazur B, Zydowicz G, Krol W. Ethanolic extract of propolis (EEP) enhances the apoptosis-inducing potential of TRAIL in cancer cells. Molecules 2009; 14(2): 738-54. google scholar
• 33. Sawicka D, Car H, Borawska MH, Niklinski J. The anticancer activity of propolis. Folia Histochem Cytobiol 2012; 50(1): 25-37. google scholar
• 34. Motomura M, Kwon KM, Suh SJ, Lee YC, Kim YK, Lee IS, et al. Propolis induces cell cycle arrest and apoptosis in human leukemic U937 cells through Bcl-2/Bax regulation. Environ Toxicol Pharmacol 2008; 26(1): 61-7. google scholar
• 35. Eom HS, Lee EJ, Yoon BS, Yoo BS. Propolis inhibits the proliferation of human leukaemia HL-60 cells by inducing apoptosis through the mitochondrial pathway. Nat Prod Res 2010; 24(4): 375-86. google scholar
• 36. Viscarra T, Buchegger K, Jofre I, Riquelme I, Zanella L, Abanto M, et al. Functional and transcriptomic characterization of carboplatin-resistant A2780 ovarian cancer cell line. Biol Res 2019; 52(1): 1-13. google scholar
• 37. Castaneda AM, MelendezCM, Uribe D, Pedroza-Dı'az J. Synergistic effects of natural compounds and conventional chemotherapeutic agents: Recent insights for the development of cancer treatment strategies. Heliyon 2022; 8(6): e09519. google scholar
• 38. Ebadi P, Fazeli M. Evaluation of the potential in vitro effects of propolis and honey on wound healing in human dermal fibroblast cells. South African J Botany 2021; 137: 414-22. google scholar
• 39. Altabbal S, Athamnah K, Rahma A, Wali AF, Eid AH, Iratni R, et al. Propolis: a detailed insight of its anticancer molecular mechanisms. Pharmaceuticals 2023; 16(3), 450. google scholar