Publication: Kronik myeloid lösemi hücrelerinde (k562) apoptotik süreç ve reaktif oksijen türleri arasındaki ilişkinin araştırılması
Abstract
1. ÖZET Amaç: Çalışmamızda kronik myeloid hücrelerini apoptoza götürebilmek ve lösemiye karşı direnç geliştirebilmek amacıyla doğal polifenollerden kuersetin ve kurkumin kullanıldı. Ancak kurkuminin biyoyararlanımının düşük olması (< %2) klinik uygulama potansiyelini sınırladığından kronik myeloid lösemi hücrelerindeki apoptotik etkisini artırmak için kurkumini kuersetin ile sinerjistik olarak kombine etmeyi denedik. Gereç ve Yöntem: WST-1 metodu ile hücre proliferasyonunun analizi yapıldı ve IC50 dozları tesbit edildi. Ardından, bu iki polifenolün sinerjik etki gösterdiği dozlar kombinasyon analiz Calcusyn) ile belirlendi. Apoptoz oranlarının tesbiti için Annexin-V, reaktif oksijen türleri oluşumunun tesbiti için DCFDA, mitokondriyal membran potansiyelinin tesbiti için JC-1 boyası kullanılarak akım sitometride analiz gerçekleştirildi. Hücreiçi glutatyon ise florometrik olarak belirlendi. Çekirdek kondensasyonu Hoechst 33342 boyası kullanılarak floresan mikroskopta gösterildi. Apoptotik mekanizmada yer alan ilgili protein ekspresyonları ise Western blot metodu ile analiz edildi. Bulgular: Kuersetin ve kurkumin kombinasyonu ile hem polifenollerin etkin dozları azaltılmış oldu hem de lösemi hücrelerinin bu sinerjistik kombinasyon ile apoptoza sürüklenmeleri artırılmış oldu. Ayrıca, bu kombinasyon kurkuminin biyoyararlanımını da artırmış oldu. Son olarak, KML hücrelerinde apoptotik süreç ve reaktif oksijen türleri arasındaki etkileşimin mekanizması aydınlatıldı. Sonuç: Kuersetin ve kurkuminin düşük konsantrasyonlarda kullanımının hem bu polifenollerin biyoyararlanımının artmasını sağlayacağını hem de kombinasyondaki sinerjistik etkileşimin kronik myeloid lösemi (KML) tedavisi için önemli olabileceğini düşünmekteyiz. Kronik myeloid lösemi (KML), kuersetin, kurkumin, apoptoz, reaktif oksijen türleri (ROT). Investigation of Relationship Between Reactive Oxygen Species and Apoptotic Process in Chronic Myeloid Leukemia (K562) Cells 2.
Aim: We have used two natural polyphenolic agents, quercetin and curcumin, to create resistance against leukemia and lead to apoptosis of cancer cells. Curcumin has limited clinical use because of its low bioavailability (< 2%). We have tried synergistic combination of curcumin with quercetin to increase its apoptotic effects on chronic myeloid leukemia cells. Methods: Cell proliferation was analyzed by WST-1 method and IC50 values were determined. Then, synergistic effects of these two polyphenols were analyzed by the Calcusyn combination analysis program. Annexin-V staining was used for detection of apoptosis, DCFDA was used for detection of reactive oxygen species and JC-1 dye was used for detection of mitochondrial membrane potential and all analyses were performed by flow cytometry. Fluorometric assaywas used tomeasureintracellular glutathione. Chromatin condensation was shown using Hoechst 33342 dye by florescence microscope. Protein expression associated with apoptotic mechanisms were analyzed using the Western blot method. Results: The rate of apoptosis was enhanced by the synergistic combination of quercetin and curcumin. The relationship between apoptotic process and reactive oxygen species formation was studied in CML cells using the combination doses. Conclusion: We suggest combined use of the two polyphenols, quercetin and curcumin to increase their bioavalibility. Low concentrations of polyphenols in combination may be useful for treatment of CML via synergistic interaction. Chronic myeloid leukemia (CML), quercetin, curcumin, apoptosis, reactive oxygen species (ROS).
Aim: We have used two natural polyphenolic agents, quercetin and curcumin, to create resistance against leukemia and lead to apoptosis of cancer cells. Curcumin has limited clinical use because of its low bioavailability (< 2%). We have tried synergistic combination of curcumin with quercetin to increase its apoptotic effects on chronic myeloid leukemia cells. Methods: Cell proliferation was analyzed by WST-1 method and IC50 values were determined. Then, synergistic effects of these two polyphenols were analyzed by the Calcusyn combination analysis program. Annexin-V staining was used for detection of apoptosis, DCFDA was used for detection of reactive oxygen species and JC-1 dye was used for detection of mitochondrial membrane potential and all analyses were performed by flow cytometry. Fluorometric assaywas used tomeasureintracellular glutathione. Chromatin condensation was shown using Hoechst 33342 dye by florescence microscope. Protein expression associated with apoptotic mechanisms were analyzed using the Western blot method. Results: The rate of apoptosis was enhanced by the synergistic combination of quercetin and curcumin. The relationship between apoptotic process and reactive oxygen species formation was studied in CML cells using the combination doses. Conclusion: We suggest combined use of the two polyphenols, quercetin and curcumin to increase their bioavalibility. Low concentrations of polyphenols in combination may be useful for treatment of CML via synergistic interaction. Chronic myeloid leukemia (CML), quercetin, curcumin, apoptosis, reactive oxygen species (ROS).