Investigation of Cytotoxic Effect of Salvia pilifera Extracts and Synthetic Chlorogenic and Caffeic Acids on DU145 Prostate Cancer Cells Line

Önder Yumrutaş; Mustafa Pehlivan; Celal Güven; İbrahim Bozgeyik; Esra Bozgeyik; Pınar Yumrutaş; Ebru Temiz; Fatih Üçkardeş

doi:10.18016/ksudobil.302249

EN TR

Investigation of Cytotoxic Effect of Salvia pilifera Extracts and Synthetic Chlorogenic and Caffeic Acids on DU145 Prostate Cancer Cells Line

Abstract

Salvia species have been used in the treatment of many diseases due to their medical effects, and the effects of these species on prostate cancer cells should be investigated in more detail. In this study, we aimed to determine anti-carcinogenic activities of dichloromethane (DCM) and methanol (MeOH) extracts of Salvia pilifera and the synthetic chloregenic (CGA) and caffeic acids (CA) on DU-145 prostate cancer cells. The cytotoxicity of extracts and synthetic compounds on cell viability of DU-145 was measured by using MTT method. Induction of apoptosis was tested by using Annexin V and 7ADD staining. DNA fragmentation was evaluated in cells. Also, transcripton levels of Bax, Caspase 3, Caspase 9 and Bcl-2 and Bcl-xL genes were determined. Lastly, the phenolic compounds in MeOH extract were determined by HPLC. In MTT test, extracts, CGA and CA were found to be diminished proliferation of DU145 cells. However, in apoptosis assay, no apoptotic activity for extract and synthetic compounds was observed. In DNA fragmentation test, while no significant difference in extracts group was observed as compared to controls, fragmentation as swab in synthetic compound groups was observed. Small changes were observed in transcription levels of apoptotic and antipoptotic genes. A total of 11 phenolic acids were determined including fumaric acid, gallic acid, gallocatechin, catechin, oleorufein, 4-hydroxybenzoic acid, caffeic acid, syringic acid, ellagic acid, 3-hydroxy cinnamic acid and protocatechuic acid. Results of the present study suggest that S. pilifera extracts and synthetic CGA and CA might have cytotoxic effects on DU145 cell at certain concentration (≥50 µg ml^-1 for DCM; ≥ 100 µg ml^-1 for MeOH; ≥ 1 µg ml^-1 for CGA and CA) yet that these effects may be manifested through another pathway but apoptosis.

Keywords

S. pilifera extract,Caffeic acid,Chlorogenic acid,Cytotoxicity,Apoptosis,Prostate cancer

Prostat Kanser Hücre Hattı DU145 Üzerine Salvia Pilifera Ekstraktları ile Sentetik Klorojenik ve Kafeik Asidin Sitotoksik ve Apoptotik Etkilerinin Incelenmesi

Öz

Salvia türleri tıbbi etkilerinden dolayı pek çok hastalığın tedavisinde kullanılmaktadır, Ancak bu türlerin prostat kanseri üzerine etkilerini gösteren çalışmaların sayısı sınırldır. Bu çalışmada, S. pilifera diklorometan (DCM) ve metanol (MeOH) ekstraktları ile sentetik klorojenik (CGA) ve kafeik asitin (CA) DU145 prostat kanser hücreleri üzerine antikanser aktivitelerinin belirlenmesi amaçlanmıştır. DU145 hücrelerinin canlılığı MTT boyama testi ile belirlenmiştir. Apoptozun indüklenmesi Annexine V ve 7ADD boyama kiti kullanılarak belirlenmiştir. Özüt uygulaması sonrasında hücrelerin DNA’ları izole edilerek fragmentasyona bakılmıştır. Son olarak, BAX, Kaspaz3 ve 9, BcL2 ve BcL-xL gen ekspresyon seviyeleri ölçülmüştür. Bununla birlikte, MeOH ektraktındaki olası fenolik bileşikler HPLC ile belirlenmiştir. MTT testinde, ekstrakt ve sentetik fenoliklerin DU145 hücrelerinin canlılığını azalttığı tespit edilmiştir. Ancak apoptoz indükleme testinde ekstrakt ve fenoliklerin herhangi bir aktivite sergilemedikleri belirlenmiştir. DNA fragmentasyon testinde, kontrolle kıyaslandığında ekstrakt uygulanan gruplarda bir farklılık görülmezken, CGA ve CA uygulama gruplarında sürüntü şeklinde parçalanma gözlenmiştir. Ayrıca, apoptotik ve antiapoptotik gen ekspresyonlarında zayıf değişimler gözlenmekle birlikte bu değişimler istatistiksel olarak önemli bulunmamıştır. Ayrıca, S. pilifera’da fumarik asit, gallik asit, gallokateşin, kateşin, oleorufein, 4-hidroksibenzoik asit, kafeik asit, sirinjik asit, ellajik asit, 3-hidroksi sinnamik asit ve protokateşik asit belirlenmiştir. Sonuç olarak, S. pilifera DCM ve MeOH özütleri ve kullanılan sentetik fenolik asitlerin prostat kanser hücreleri üzerine belirli dozlarda (DCM için ≥50 µg ml^-1; MeOH için ≥ 100 µg ml^-1; CGA and CA için ≥ 1 µg ml^-1) sitotoksit etki gösterdikleri, ancak bu etkinin apoptoz yolağıyla ilişkili olmadığı belirlenmiştir.

Anahtar Kelimeler

S. pilifera ekstrakt,Kafeik asit,Klorojenik asit,Apoptosis,Prostat kanser

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Details

Primary Language

English

Subjects

-

Journal Section

Research Article

Authors

Önder Yumrutaş ^*

Mustafa Pehlivan

Celal Güven

İbrahim Bozgeyik

Esra Bozgeyik

Pınar Yumrutaş

Ebru Temiz

Fatih Üçkardeş

Publication Date

April 30, 2018

Submission Date

March 29, 2017

Acceptance Date

May 22, 2007

Published in Issue

Year 2018 Volume: 21 Number: 2

DOI

https://doi.org/10.18016/ksudobil.302249

IZ

https://izlik.org/JA67UK52TR

Cite

RIS / Bibtex

APA

Yumrutaş, Ö., Pehlivan, M., Güven, C., Bozgeyik, İ., Bozgeyik, E., Yumrutaş, P., Temiz, E., & Üçkardeş, F. (2018). Investigation of Cytotoxic Effect of Salvia pilifera Extracts and Synthetic Chlorogenic and Caffeic Acids on DU145 Prostate Cancer Cells Line. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, 21(2), 141-147. https://doi.org/10.18016/ksudobil.302249

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