TY - JOUR T1 - Propolis: Intrinsic Pathway-Induced Apoptosis, G1 Cell Cycle Arrest, Reduced Chemotherapeutic Resistance in Adenocarcinoma, and Healthy Cell Preservation AU - Kervancıoğlu Demirci, Elif AU - İşçi, Nursel AU - Doğan, Sibel AU - Yardım, Sahika AU - Solakoğlu, Seyhun AU - Tahralı, İlhan AU - Deniz, Günnur AU - Bekiroğlu, Gülnaz Nural PY - 2023 DA - December DO - 10.26650/experimed.1343683 JF - Experimed PB - Istanbul University WT - DergiPark SN - 2667-5846 SP - 194 EP - 204 VL - 13 IS - 3 LA - en AB - 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. KW - 3T3 KW - apoptosis KW - electron microscopy KW - endometrium KW - Ishikawa KW - migrasome CR - 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 CR - 2. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin 2016; 66(1): 7-30. google scholar CR - 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 CR - 4. Nishida M. The Ishikawa cells from birth to the present. Hum Cell 2002; 15(3): 104-17. google scholar CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 10. Castaldo S, Capasso F. Propolis, an old remedy used in modern medicine. Fitoterapia 2002; 73 Suppl 1:S1-6. google scholar CR - 11. Burdock GA. Review of the biological properties and toxicity of bee propolis (propolis). Food Chem Toxicol 1998; 36(4): 347-63. google scholar CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 22. Kelland L The resurgence of platinum-based cancer chemotherapy Nat Rev Cancer 2007; 7(8): 573-84 google scholar CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 33. Sawicka D, Car H, Borawska MH, Niklinski J. The anticancer activity of propolis. Folia Histochem Cytobiol 2012; 50(1): 25-37. google scholar CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 CR - 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 UR - https://doi.org/10.26650/experimed.1343683 L1 - https://dergipark.org.tr/en/download/article-file/3334726 ER -