Yumurtalık kanseri hücre hatları üzerinde hidroksitirozolün sitotoksik ve apoptotik etkileri: sitotoksik etkinin ERK 1/2 yolağı ile ilişkisi

Amaç: Hiroksitirozolün (HT) çeşitli kanser hücre hatları üzerindeki antitümoral, çoğalmayı inhibe etme ve apoptotik etkileri olduğu gösterilmiştir. Bu çalışmada, insan yumurtalık kanser hücre hatları OVCAR-3 ve MDAH-2774 üzerinde HT'nin sitotoksik ve apoptotik etkilerini ve onun sitotoksik etkisinin fosfotidil 3-kinaz/Akt (PI3K/Akt) ve hücre dışı sinyalle düzenlenen kinaz 1/2 (ERK 1/2) ile ilişkisini araştırdık. Yöntem: HT'nin, 24, 48, 72 ve 96 saatlik OVCAR-3 ve MDAH-2774 hücre kültürleri üzerinde sitotoksik ve apoptotik etkilerini belirlemek için, XTT hücre canlılık testi, hücre ölümünü belirleyen ELISA plus ve insan apoptoz array testi kullanıldı. HT'nin sitotoksik etkisinin PI3K/Akt ve ERK 1/2 sinyal yolakları ile ilişkisi, yolakların spesifik inhibitörleri kullanılarak araştırıldı. Bulgular: HT'nin IC50 değeri MDAH-2774 hücrelerinde 72. saatte 102.3 ?M, OVCAR-3 hücrelerinde 96. saatte 51.5 ?M bulundu. En yüksek apoptotik etki OVCAR-3 için 96. saatte 100 ?M ve MDAH-2774 için 72. saatte 50 ?M'da görüldü. HT, MDAH-2774 hücrelerinde Bad, Bax, aktif kaspaz-3, Htra2/Omi'nin dahil olduğu pro-apoptotik moleküllerin seviyesini ve TRAIL R1/DR4, TRAIL R2/DR5, FAS/TNFRSF6 içeren pro-apoptotik ölüm reseptörlerinin seviyesini artırdı. OVCAR-3 hücrelerinde HT, Bcl-2, pro-caspase 3 and CIAP-1, CIAP-2, XIAP, Livin, Survivin seviyelerini azalttı ve sitokrom-c seviyesini arttırdı. Hücre canlılığı, HT ile FR180204 uygulamasının sonucunda her iki hücre hattında arttı. Sonuç: Bu sonuçlar HT'nin OVCAR-3 ve MDAH-2774 hücreleri üzerinde sitotoksik ve apoptotik etki potansiyeline sahip olduğunu göstermektedir. Her iki kültür hücrelerinde HT'nin sitotoksik etkisi ERK 1/2 sinyal yolağını inhibe ederek gerçekleşiyor olabilir.

Cytotoxic and apoptotic effects of hydroxytyrosol on ovarian cancer cell lines: relationship of cytotoxic effect with ERK 1/2 pathway

Objective: It has been shown that hydroxytryrosol (HT) has antitumoral, anti-proliferative and apoptotic effects on various cancer cell line types. In this study, we investigated cytotoxic and apoptotic effects of HTon human ovarian cancer cell lines OVCAR-3 and MDAH-2774 and relationship of its cytotoxic effects with phosphatidylinositol 3-kinase/Akt (PI3K/Akt) and extracellular signal-regulated kinase 1/2 (ERK 1/2) signaling pathways. Method: XTT cell proliferation kit, Cell Death Detection Elisa Plus Kit and Human Apoptosis Array test were used to determine the cytotoxic and apoptotic effects of HT in OVCAR-3 and MDAH-2774 cell lines at 24, 48, 72 and 96 hours (h). Effect of HT on PI3K/Akt and ERK 1/2 signaling pathways were investigated by using specific inhibitors of these pathways. Results: IC50 values of HT were found to be 102.3 ?M in MDAH-2774 cells at 72 h and 51.5 ?M in OVCAR-3 cells at 96 h. Highest apoptotic effect was seen at 100 ?M concentration in OVCAR-3 cells at 96 h and at 50 ?M at 72 h in MDAH-2774. HT increased the levels of pro-apoptotic molecules including Bad, Bax, active caspase-3, Htra2/Omi and of proapoptotic death receptors TRAIL R1/DR4, TRAIL R2/DR5, FAS/TNFRSF6 and decreased Survivin levels in MDAH-2774 cells. In OVCAR-3 cells, HT decreased Bcl-2, pro-caspase 3 and CIAP-1, CIAP-2, XIAP, Livin, Survivin levels and increased cytochrome-c level. The cell vitality was increased in both of the cell lines as the result of the treatment with HT and FR180204. Conclusions: These results show that HT has potential cytotoxic and apoptotic effect on OVCAR3 and MDAH-2774 cells. In both cell lines, the cytotoxic effect of HT could be ocur inhibiting through ERK 1/2 signaling pathway.

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1. Ali AY, Farrand L, Kim JY, Byun S, Suh JY, Lee HJ, Tsang BK. Molecular determinants of ovarian cancer chemoresistance: new insights into an old conundrum. Ann N Y Acad Sci 2012; 1271: 58-67.
2. Smolle E, Taucher V, Pichler M, Petru E, Lax S, Haybaeck J. Targeting Signaling Pathways in Epithelial Ovarian Cancer. Int J Mol Sci 2013; 14: 9536-55.
3. Longuespée R, Boyon C, Desmons A, Vinatier D, Leblanc E, Farré I, Wisztorski M, Ly K,D'Anjou F, Day R, Fournier I, Salzet M. Ovarian cancer molecular pathology. Cancer and Metastasis Rev 2012; 31: 713- 32.
4. Romero I, Bast RC. Minireview: Human Ovarian Cancer: Biology, Current Management, and Paths to Personalizing Therapy Endocrinology 2012; 153: 1593- 1602.
5. Dupont J, Reverchon M, Cloix L, Froment P, Ramé C. Involvement of adipokines, AMPK, PI3K andthe PPAR signaling pathways in ovarian follicle development and cancer. Int J Dev Biol 2012; 56: 959-67.
6. Bernini R, Merendino N, Romani A, Velotti F. Naturally occurring hydroxytyrosol: synthesis and anticancer potential. Curr Med Chem 2013; 20: 655-70.
7. Granados-Principal S, Quiles J, Ramirez-Tortosa CL, SanchezRovira P, Ramirez-Tortosa MC. Hydroxytyrosol: from laboratory investigations to future clinical trials. Nutr Rev 2010; 68: 191-206.
8. Guichard C, Pedruzzi E, Fay M, Marie JC, Braut-Boucher F, Daniel F, Grodet A, Gougerot-Pocidalo MA, Chastre E, Kotelevets L, Lizard G, Vandewalle A, Driss F, Ogier-Denis E. Dihydroxyphenylethanol induces apoptosis by activating serine/threonine protein phosphatase PP2A and promotes the endoplasmic reticulum stres response in human colon carcinoma cells. Carcinogenesis 2006; 27: 1812-27.
9. Corona G, Deiana M, Incani A, Vauzour D, Dessì MA, Spencer JP. Inhibition of p38/CREB phosphorylation and COX-2 expression by olive oil polyphenols underlies their anti-proliferative effects. Biochem Biophys Res Commun 2007; 362: 606-11.
10. Fabiani R, Rosignoli P, De Bartolomeo A, Fuccelli R, Morozzi G. Inhibition of cell cycle progression by hydroxytyrosol is associated with upregulation of cyclin-dependent protein kinase inhibitors p21(WAF1/Cip1) and p27(Kip1) and with induction of differentiation in HL60 cells. J Nutr 2008; 138: 42-8.
11. Corona G, Deiana M, Incani A, Vauzour D, Dessi MA, Spencer JPE. Hydroxytyrosol inhibits the proliferation of human colon adenocarcinoma cells through inhibition of ERK1/2 and cyclin D1. Mol Nutr Food Res 2009; 53: 897- 903.
12. Bouallagui Z, Han J, Isoda H, Sayadi S. Hydroxytyrosol rich extract from olive leaves modulates cell cycle progression in MCF-7 human breast cancer cells. Food Chem Toxicol 2011; 49: 179-84.
13. Notarnicola M, Pisanti S, Tutino V, Bocale D, Rotelli MT, Gentile A, Memeo V, Bifulco M, Perri E, Caruso MG. Effects of olive oil polyphenols on fatty acid synthase gene expression and activity in human colorectal cancer cells. Genes Nutr 2011; 6: 63-9.
14. Tuğrul B, Oktay LM. The role of hydroxytyrosol in health and disease. Turkish Journal of Oncology 2014; 29: 166-72.
15. Nan JN, Ververis K, Bollu S, Rodd AL, Swarup O, Karagiannis TC. Biological effects of the olive polyphenol, hydroxytyrosol: An extra view from genome-wide transcriptome analysis. Hell J Nucl Med 2014; 17: 62-9.
16. Han J, Talorete TPN, Yamada P, Isoda H. Anti-proliferative and apoptotic effects of oleuropein and hydroxytyrosol on human breast cancer MCF-7 cells. Cytotechnology 2009; 59: 45-53.
17. Li S, Han Z, Ma Y, Song R, Pei T, Zheng T, Wang J, Xu D, Fang X, Jiang H, Liu L. Hydroxytyrosol inhibits cholangiocarcinoma tumor growth: an in vivo and in vitro study. Oncol Rep 2014; 31: 145-52.
18. Fabiani R, Sepporta MV, Patrizia R, De Bartolomeo A, Crescimanno M, Morozzi G. Anti proliferative and Pro-apoptotic activities of Hydroxytyrosol on Different Tumour Cells: The Role Extracellular production of Hydrogen Peroxide. Eur J Nutr 2012; 51: 455-64.
19. Fabiani R, De Bartolomeo A, Rosignoli P, Servili M, Montedoro GF, Morozzi G. Cancer chemoprevention by hydroxytyrosol isolated from virgin olive oil through G1 cell cycle arrest and apoptosis. Eur J Cancer Prev 2002; 11: 351-58.
20. D'Angelo S, Ingrosso D, Migliardi V, Sorrentino A, Donnarumma G, Baroni A, Masella L, Tufano MA, Zappia M, Galletti P. Hydroxytyrosol, a natura lantioxidant from olive oil, prevents protein damage induced by longwave ultraviolet radiation in melanoma cells. Free Radic Biol Med 2005; 38: 908-19.
21. Sirianni R, Chimento A, De Luca A, Casaburi I, Rizza P, Onofrio A, Iacopetta D, Puoci F, Ando S, Maggiolini M, Pezzi V. Oleuropein and hydroxytyrosol inhibit MCF-7 breast cancer cell proliferation interfering with ERK 1/2 activation. Mol Nutr Food Res 2010; 54: 833- 40.