Prostat Kanser Hücre Hattı DU145 Üzerine Salvia Pilifera Ekstraktları ile Sentetik Klorojenik ve Kafeik Asidin Sitotoksik ve Apoptotik Etkilerinin Incelenmesi
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.
Investigation of Cytotoxic Effect of Salvia pilifera Extracts and Synthetic Chlorogenic and Caffeic Acids on DU145 Prostate Cancer Cells Line
Salvia species have been used in the treatment of many diseases dueto their medical effects, and the effects of these species on prostatecancer cells should be investigated in more detail. In this study, weaimed to determine anti-carcinogenic activities of dichloromethane(DCM) and methanol (MeOH) extracts of Salvia pilifera and thesynthetic chloregenic (CGA) and caffeic acids (CA) on DU-145prostate cancer cells. The cytotoxicity of extracts and syntheticcompounds on cell viability of DU-145 was measured by using MTTmethod. Induction of apoptosis was tested by using Annexin V and7ADD staining. DNA fragmentation was evaluated in cells. Also,transcripton levels of Bax, Caspase 3, Caspase 9 and Bcl-2 and BclxLgenes were determined. Lastly, the phenolic compounds inMeOH extract were determined by HPLC. In MTT test, extracts,CGA and CA were found to be diminished proliferation of DU145cells. However, in apoptosis assay, no apoptotic activity for extractand synthetic compounds was observed. In DNA fragmentation test,while no significant difference in extracts group was observed ascompared to controls, fragmentation as swab in synthetic compoundgroups was observed. Small changes were observed in transcriptionlevels of apoptotic and antipoptotic genes. A total of 11 phenolicacids were determined including fumaric acid, gallic acid,gallocatechin, catechin, oleorufein, 4-hydroxybenzoic acid, caffeicacid, syringic acid, ellagic acid, 3-hydroxy cinnamic acid andprotocatechuic acid. Results of the present study suggest that S.pilifera extracts and synthetic CGA and CA might have cytotoxiceffects 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 theseeffects may be manifested through another pathway but apoptosis.
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