Kuersetinin İnsan Meme Kanseri Hücre Hattı Üzerine Etkisinin İncelenmesi

Yıl 2022, Cilt: 6 Sayı: 3, 261 - 269, 25.12.2022

Münevver Baran , Özge Göktepe , Gözde Özge Önder , Zeynep Burcin Gonen , Arzu Yay

https://doi.org/10.46332/aemj.1005558

Öz

Amaç: Meme kanseri, farklı epigenetik değişiklikler ve genetik mutasyonlar nedeniyle ortaya çıkan yaygın bir kanserdir. Doğal olarak oluşan Kuersetin, antioksidan, anti-inflamatuar ve anti-kanser özellikler sergileyen ve çok sayıda hücre içi hedefi yöneten bir flavonoid’dir. Bu çalışma da, MCF-7 meme kanseri hücre hattında Kuersetinin apopitoz /otofaji süreçleri üzerine olası etkisi-nin incelenmesi amaçlandı.

Araçlar ve Yöntem: Kuersetinin IC50 değerini 3-4.5-dimetil-tiyazolil–2.5-difeniletrazolyum bromid (MTT) analiz ile belirlemek için MCF-7 meme kanseri hücre hattına farklı konsantrasyonlar da ve sürelerde (24, 48 ve 72. saatte) kuersetin uygulandı ve gruplar oluşturuldu. Kontrol grubu; hiç bir uygulama yapılmayan grup, Kuersetin 1; kuersetinin IC50 dozunun uygulandığı grup, Kuersetin 2; kuersetin’in IC50 dozunun yarısının uygulandığı grup, D 1; kuersetin 1 grubunda kuersetin hazırlanmasında kullanı-lan DMSO oranının uygulandığı grup ve D 2; kuersetin 2 grubun da kuersetin hazırlanmasında kullanılan DMSO oranının uygu-landığı grup. Kuersetinin apopitotik etkisini değerlendirmek için TUNEL boyama yöntemi kullanılırken, otofajik etkisini ortaya çıkarmak için LC3/2 ve Beklin-1 immünofloresan boyama yöntemi uygulandı.

Bulgular: Kuersetinin hücreler üzerine etkisine bakıldığında, hücre canlılığının doz ve zamana bağlı olarak inhibe olduğu görüldü. Bunun yanında otofaji belirteçleri olan Beklin-1 ve LC3/2 immunre aktivitesinin de arttığı gözlendi.

Sonuç: Sonuç olarak, kuersetinin apopitoz ve otofaji yolakları ile MCF-7 meme kanseri hücrelerini ölüme götürdüğü gözlendi.

Anahtar Kelimeler

Apoptoz, Kuersetin, MCF-7 hücre hattı, Otofaji

Investigation of the Effect of Quercetin on Human Breast Cancer Cell Line

Öz

Purpose: Breast cancer is a common cancer that occurs due to different epigenetic changes and genetic mutations. Naturally occurring Quercetin is a flavonoid that exhibits antioxidant, anti-inflammatory and anti-cancer properties and manages multiple intracellular targets. In this study, it was aimed to investigate the possible effect of Quercetin on apoptosis/autophagy processes in MCF-7 breast cancer cell line.

Materials and Methods: Quercetin was applied to MCF-7 breast cancer cell line at different concentrations and times (24, 48 and 72 hours) to determine the IC50 value of quercetin by 3-4.5-dimetil-tiyazolil–2.5-difeniletrazolyum bromid (MTT) analysis, and groups were formed. Control group; no treatment group, Quercetin 1; IC50 dose of quercetin administered group, Quercetin 2; group in which half the IC50 dose of quercetin was administered, D 1; In the quercetin 1 group, the DMSO ratio used in the preparation of the quercetin was applied, and D 2; In the quercetin 2 group, the DMSO ratio used in the preparation of the quer-cetin was applied, While TUNEL staining method was used to evaluate the apoptotic effect of quercetin, LC3/2 and Beclin-1 immunofluorescence staining method was applied to reveal its autophagic effect.

Results: When the effect of quercetin on cells was examined, it was observed that cell viability was inhibited depending on dose and time. In addition, it was observed that the autophagy markers Beclin-1and LC3/2 immunoreactivity increased, too.

Conclusion: In conclusion, it was observed that quercetin led to death of MCF-7 breast cancer cells through apoptosis and autophagy pathways.

Anahtar Kelimeler

Apoptosis, Autophagy, MCF-7 cell line, Quercetin

Kaynakça

• 1. Akkuzu MZ, Küçüköner M, Irtegun S, ve ark. Meme kanserinde Brca-1 ve Brca-2’de sık görülen polimorfizm mutasyonların bölgemizde varlığı. Dicle Med J. 2019;46(4):623-631.
• 2. Kampa M, Nifli AP, Notas G, Castanas E. Polyphenols and cancer cell growth. Rev. Physiol. Biochem. Pharmacol, 2007;159:79-113.
• 3. Sonnenblick A, Fumagalli D, Sotiriou C, Piccart M. Is the differentiation into molecular subtypes of breast cancer important for staging, local and systemic therapy, and follow up? Cancer Treat Rev. 2014;40:1089-1095.
• 4. Eliyatkin N, Yalcin E, Zengel B, Aktaş S, Vardar E. Molecular classification of breast carcinoma: from traditional, oldfashioned way to a new age, and a new way. J Breast Heal. 2015;11:59-66.
• 5. Surh YJ. Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer. 2003;3(10):768-780.
• 6. Thomasset SC, Berry DP, Garcea G, Marczylo T, Steward WP, Gescher AJ. Dietary polyphenolic phytochemicalspromising cancer chemopreventive agents in humans? A review of their clinical properties. Int J Cancer. 2007;120(3):451-458.
• 7. Birman, H. Bitkisel flavonoid bileşiklerinin biyoaktiviteleri ve muhtemel etki mekanizmaları. İst Tıp Fak Derg. 2012;75(3):46-49.
• 8. Cárdenas M, Marder M, Blank VC, Roguin LP. Antitumor activity of some natural flavonoids and synthetic derivatives on various human and murine cancer cell lines. Bioorg Med Chem. 2006;14(9):2966-2971.
• 9. Russo M, Spagnuolo C, Tedesco I, Bioltto S, Russo GL. The flavonoid quercetin in disease prevention and therapy: Facts and fancies. Biochem Pharmacol. 2012;83(1):6-15.
• 10. Chien SY, Wu YC, Chung JG. Quercetin-induced apoptosis acts through mitochondrial and caspase-3-dependent pathways in human breast cancer MDA-MB-231 cells. Hum Exp Toxicol. 2009;28(8):493-503.
• 11. Lamson DW, Brignall MS. Antioxidants and cancer III: Quercetin. Altern Med Rev. 2000;5(3):196-208.
• 12. Comşa Ş, Cîmpean AM, Raica M. The story of MCF-7 breast cancer cell line: 40 Years of experience in research. Anti-cancer Res. 2015;35(6):3147-3154.
• 13. Sweeney EE, McDaniel RE, Maximov PY, Fan P, Craig Jordan V. Models and mechanisms of acquired antihormone resistance in breast cancer: Significant clinical progress despite limitations. Horm. Mol. Biol. Clin. Investig. 2012;9(2):143-163.
• 14. Granato M, Rizzello C, Gilardini Montani MS, et al. Quercetin induces apoptosis and autophagy in primary effusion lymphoma cells by inhibiting PI3K/AKT/mTOR and STAT3 signaling pathways. J Nutr. Biochem. 2017;41:124-136.
• 15. Jia L, Huang S, Yin X, Zan Y, Guo Y, Han L. Quercetin suppresses the mobility of breast cancer by suppressing glycolysis through Akt-mTOR pathway mediated autophagy induction. Life Sci. 2018;208:123-130.
• 16. Hashemzaei M, Delarami Far A, Yari A, et al. Anticancer and apoptosis inducing effects of quercetin in vitro and in vivo. Oncol Rep. 2017;38(2):819-828.
• 17. Kim H, Moon JY, Ahn KS, Cho SK. Quercetin induces mitochondrial mediated apoptosis and protective autophagy in human glioblastoma U373MG cells. Oxid Med Cell Longev. 2013;2013:596496.
• 18. Liu Y, Gong W, Yang ZY, et al. Quercetin induces protective autophagy and apoptosis through ER stress via the p-STAT3/Bcl-2 axis in ovarian cancer. Apoptosis. 2017;22(4):544-557.
• 19. Mosmann T. Rapid colorimetric assay for celluler growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1-2):55-63.
• 20. Hashemzaei M, Delarami Far A, Yari, A, et al. Anticancer and apoptosis inducing effects of quercetin in vitro and in vivo. Oncology reports. 2017;38(2):819-828.
• 21. Corre LL, Chalabi N, Delort L, Bignon Y, Bernard-Gallon DJ. Differential expression of genes induced by resveratrol in human breast cancer cell lines. Nutr Cancer. 2006;56(2):193-203.
• 22. Kandaswami C, Lee LT, Lee PP, et al. The antitumor activities of flavonoids. In Vivo. 2005;19(5):895-909.
• 23. Maggiolini M, Bonofiglio D, Marsico S, et al. Estrogen receptor α mediates the proliferative but not the cytotoxic dose-dependent effects of two major phytoestrogens on human breast cancer cells. Mol Pharmacol. 2001;60(3):595-602.
• 24. Wang L, Lee IM, Zhang SM, Blumberg JB, Buring JE, Sesso HD. Dietary intake of selected flavonols, flavones, and flavonoidrich foods and risk of cancer in middleaged and older women. Am. J. Clin. Nutr. 2009;89(3):905-912.
• 25. Lautraite S, Musonda AC, Doehmer J, EdwardsGO, Chipman JK. Flavonoids inhibit genetic toxicity produced by carcinogens in cells expressing CYP1A2 and CYP1A1. Mutagenesis. 2002;17(1):45-53.
• 26. Haghiac M, Walle T. Quercetin induces necrosis and apoptosis in SCC-9 oral cancer cells. Nutr Cancer. 2005;53(2):220-231.
• 27. Mozhgan FS, Seyed BJ, Bahareh H. The Cuscuta kotschyana effects on breast cancer cells line MCF7. J Med Plants Res. 2011;5(27):6344-6351.
• 28. Russo M, Spagnuolo C, Bilotto S, Tedesco I, Maiani G, Russo GL. Inhibition of protein kinase CK2 by quercetin en-hances CD95-mediated apoptosis in a human thymus-derived T cell line. Food Res Int. 2014;63(Part B):244-51.
• 29. Ranganathan S, Halagowder D, Sivasithambaram ND. Quercetin suppresses twist to induce apoptosis in MCF-7 breast cancer cells. PLoS One. 2015;10:e0141370.
• 30. Dechsupa S, Kothan S, Vergote J, et al. Quercetin, Siamois 1 and Siamois 2 induce apoptosis in human breast cancer MDA-MB-435 cells xenograft in vivo. Cancer Biol Ther. 2007:6(1);56-61.
• 31. Ishikawa Y, Kitamura M. Anti-apoptotic effect of quercetin: intervention in the JNK-and ERK-mediated apoptotic pathways. Kidney Int. 2000;58(3):1078-1087.
• 32. Chou CC, Yang JS, Lu HF, et al. Quercetin mediated cell cycle arrest and apoptosis involving activation of a caspase cascade through the mitochondrial pathway in human breast cancer MCF-7 cells. Arch Pharm Res. 2010;33(8):1181-1191.
• 33. Lee YK, Park S Y, Kim YM, Lee WS, Park OJ. Regulation of MCF-7 cell apoptosis by phytochemical Quercetin through AMPK-mTOR signaling pathway. Cancer Prev. Res. 2010;15(4):320-325.
• 34. Choi EJ, Bae SM, Ahn WS. Antiproliferative effects of quercetin through cell cycle arrest and apoptosis in human breast cancer MDA-MB-453 cells. Arch Pharm Res. 2008;31(10):1281-1285.
• 35. De Blas E, Estan MC, de Frutos, MDCG, Ramos J, del Carmen Boyano-Adanez M, Aller P. Selected polyphenols po-tentiate the apoptotic efficacy of glycolytic inhibitors in human acute myeloid leukemia cell lines. Regulation by protein kinase activities. Cancer Cell Int. 2016;16(1):1-16.
• 36. Ohsumi Y. Historical landmarks of autophagy research. Cell Res. 2014;24(1):9-23.
• 37. Nencioni A, Cea M, Montecucco F, et al. Autophagy in blood cancers: biological role and therapeutic implications. Haematologica. 2013;98(9):1335-1343.
• 38. Xie Z, Klionsky DJ. Autophagosome formation: core machinery and adaptations. Nat Cell Biol. 2007;9(10): 1102-1129.
• 39. Martinet W, De Meyer GR, Andries L, Herman AG, Kockx MM. Detection of autophagy in tissue by standard ımmunohistochemistry possibilities and limitations. Autophagy. 2006;2(1):55-57.
• 40. Rezabakhsh A, Rahbarghazi R, Malekinejad H, Fathi F, Montaseri A, Garjani A. Quercetin alleviates high glucoseinduced damage on human umbilical vein endothelial cells by promoting autophagy. Phytomedicine. 2019;56:183-193.
• 41. Wu L, Li J, Liu T, et al. Quercetin shows anti‐tumor effect in hepatocellular carcinoma LM3 cells by abrogating JAK2/STAT3 signaling pathway. Cancer Med. 2019;8(10):4806-4820.