CYTOTOXIC EFFECTS OF 3,4-DIHYDROXYPHENYL ETHANOL, AN OLIVE MILL WASTE WATER PHENOLIC, ON DIFFERENT PROSTATE CANCER CELL LINES

Abstract

The rich phenolic content of olive mill waste water obtained as by-product during olive oil production and its antioxidant, antithrombotic, anti-inflammatory, hypocholesterolemic, antimicrobial, antiviral, and anticancerogenic activities have been revealed by scientific studies. One of the main components of olive oil is 3,4-dihydroxyphenyl ethanol (3,4-DHPEA). Although the effect of 3,4-DHPEA was studied on many types of cancer, studies on prostate cancer are limited in the literature. For this purpose, similar and different cytotoxic effects of 3,4-DHPEA on 5 human prostate cancer cell lines differing in origin, genetic structure, and characteristic features were investigated. The 3,4-DHPEA was applied on prostate cancer cell lines (LNCaP, C4-2, 22Rv1, PC3, and DU-145) at different concentrations for up to 72 hours and cell viabilities were measured by the Cell Titer-Glo luminescence assay. All experiments were performed in 3 parallel and repeated 3 times. Results were compared with analysis of variance (ANOVA) and the probabilities less than 5% (P<0.05) were evaluated as significant. The 3,4-DHPEA showed time- and dose-dependent cytotoxicity on all cell lines used. The most sensitive cell line was LNCaP with 72 hr IC50=25.64 µg/mL value and the least sensitive cell line was PC3 with 72 hr IC50=58.81 µg/mL value. Nearly entire LNCaP population was found to be dead after 75 μg/mL of 3,4-DHPEA treatment for 48 h. The fact that different cancer cell lines developed from prostate tissue show different degrees of sensitivity to 3,4-DHPEA could be due to the different origins, different genetic structures (e.g. some cells have phosphatase and tensin homolog (PTEN) gene mutation), and different characteristic features (e.g. some of the cell lines used were androgen-dependent and the others are androgen-independent). 

Keywords

• Prostate cancer
• 3-4-DHPEA
• Cytotoxicity
• Olive mill waste water

ZEYTİN KARASUYU FENOLİKLERİNDEN 3,4-DİHİDROKSİFENİL ETANOLÜN FARKLI PROSTAT KANSERİ HÜCRE HATLARINDAKİ SİTOTOKSİK ETKİLERİ

Öz

Zeytinyağı üretimi sırasında oluşan yan ürünlerden zeytin karasuyunun zengin fenolik içeriği ve buna bağlı olarak antioksidan, antitrombotik, antiinflamasyon, hipokolesterolemik, antimikrobiyal, antiviral ve antikanserojenik aktiviteleri bilimsel çalışmalarla ortaya konmuştur. Zeytin karasuyunun temel bileşenlerinden biri 3,4-dihidroksifenil etanol (3,4-DHPEA)’dür. 3,4-DHPEA’nın birçok kanser tipi üzerinde antikanserojenik etkileri araştırılmasına rağmen prostat kanseri üzerindeki etkilerini irdeleyen çalışmalar literatürde sınırlı sayıdadır. Bu amaçla çalışmamızda, 3,4-DHPEA’nın orijin, genetik yapı ve karakteristik özellikler bakımından farklı 5 insan prostat kanseri hücre hatları üzerindeki benzer ve farklı sitotoksik etkileri araştırılmıştır. 3,4-DHPEA farklı konsantrasyonlarda prostat kanseri hücre hatları (LNCaP, C4-2, 22Rv1, PC3 ve DU-145) üzerinde 72 saate kadar uygulandı ve hücre canlılıkları Cell Titer-Glo luminesan testi uygulanarak belirlendi. Tüm deneyler 3 paralel yapıldı ve 3 kere tekrarlandı. Sonuçlar varyans analizi (ANOVA) ile karşılaştırıldı ve %5’den düşük olasılıklar (P<0.05) anlamlı olarak değerlendirildi. 3,4-DHPEA, kullanılan tüm hücre hatları üzerinde zamana ve doza bağlı sitotoksisite göstermiştir. En hassas hücre hattının 72 saatlik IC50=25.64 µg/mL değeriyle LNCaP ve en az hassas hücre hattının ise 72 saatlik IC50=58.81 µg/mL değeriyle PC3 olduğu tespit edildi. LNCaP popülasyonunun 48 saat 75 µg/mL 3,4-DHPEA muamelesi sonunda tamamına yakınının öldüğü görülmüştür. Prostat dokusundan gelişen farklı kanser hücre hatlarının 3,4-DHPEA’ya karşı farklı derecelerde hassasiyet göstermeleri hücrelerin farklı orijin, farklı genetik yapı (bazı hücrelerin fosfataz ve tensin homolog (PTEN) gen mutasyonuna sahip olması gibi) ve farklı karakteristik özelliklere (kullanılan hücre hatlarının bazılarının androjene bağımlı bazılarının androjenden bağımsız olması gibi) sahip olması sebebiyle olduğu düşünülmüştür. 

Anahtar Kelimeler

• Prostat kanseri
• 3-4-DHPEA
• Sitotoksisite
• Zeytin karasuyu

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