Epigallokateşin-3- gallat uygulamasına bağlı olarak kronik miyeloid lösemi hücrelerinde genetik ve epigenetik otofaji regülatörlerinin ekspresyon değişimlerinin değerlendirilmesi

Abstract

Amaç: Kronik miyeloid lösemi, Bcr-Abl1 füzyon tirozin kinazının oluşumu aktivitesi ile tanımlanır. Füzyon proteinini hedefleyen inhibitörün keşfi sağ kalım oranlarında büyük artış meydana getirmiştir. Ancak bu inhibitöre karşı direnç gelişimi yeni terapi hedeflerinin belirlenmesini zorunlu kılmaktadır. Otofaji, güncel kanser araştırmalarında dikkat çekici bir iki yönlü hedeftir. Kanser hücrelerini stres koşullarına adapte etmeyi sağladığı için genellikle otofajinin inhibisyou hedeflenmektedir. Epigallokateşin-3-gallat yeşil çayda bulunan temel bir fitokimyasaldır. mikroRNA’lar gibi epigenetik regülatörlerin üzerindeki etkinliği tedavide olduğu kadar hastaların beslenme önerilerinin düzenlenmesinde de önem taşımaktadır. Bu çalışmada bir flavanoid olan epigallokateşin-3-gallatın kronik miyeloid lösemi hücrelerinde otofajiyi düzenleyen genetik ve epigenetik faktörler üzerindeki etkinliğinin araştırılması amaçlanmıştır.

Gereç ve yöntem: mikroRNA-mRNA etkileşimini değerlendiren veritabanları kullanılarak otofaji ilişkili genleri hedefleyen mikroRNA’lar belirlenmiştir. 50 µikromolar Epigallokateşin-3-gallat uygulanan K-562 hücrelerinde gerçek zamanlı kantitatif PCR yöntemi kullanılarak otofaji ilişkili genlerin ve bu genleri hedefleyen mikroRNA’ların ekspresyon seviyeleri belirlenmiştir.

Bulgular: Epigallokateşin-3-gallat uygulaması K-562 hücrelerinde otofajinin pozitif regülatörü olan genlerin ekspresyon seviyesinde anlamlı azalışa, bu genleri hedefleyen mikroRNA’ların ekspresyon seviyelerinde ise anlamlı artışa neden olmuştur.

Sonuç: Elde edilen veriler doğrultusunda epigallokateşin-3-gallatın, kronik miyeloid lösemi hücrelerinde genetik ve epigenetik regülasyonla otofaji inhibisyonuna neden olduğu belirlenmiştir. Otofajinin çift yönlü mekanizması göz önünde bulundurulduğunda bu sonuç epigallokateşin-3-gallatın ilaç potansiyeli olma konusunda olduğu kadar hastaların beslenme şeklinde de önem taşıyan bir kimyasal olduğu gerçeğini ortaya koymaktadır.

Keywords

• Kronik Miyeloid Lösemi,miRNA,Otofaji

Evaluation of expression changes of genetic and epigenetic autophagy regulators in chronic myeloid leukemia cells due to epigallocatechin-3-gallate treatment

Abstract

Purpose: Chronic myeloid leukemia is characterized by the formation and activation of Bcr-Abl1 fusion tyrosine kinase. Discovery of the inhibitor which targets the fusion protein results in a significant increase in survival rates.  However the development of resistance to the inhibitor necessitates determining new therapy targets. Autophagy is a remarkable dual target in recent cancer researches. It is usually aimed at inhibition of autophagy because autophagy allows cells to adapt to stress conditions. Epigallocatechin-3-gallate is a basic phytochemical found in green tea. The efficacy of epigenetic regulators, such as microRNAs, is important in the regulation of dietary recommendations as well as in treatment. In this study, it was aimed to investigate the effect of epigallocatechin-3-gallate, a flavanoid, on genetic and epigenetic factors regulating autophagy in chronic myeloid leukemia cells.

Materials and methods: microRNAs which are target autophagy-related genes have been identified using appropriate databases. Expression levels of autophagy-related genes and microRNAs targeting these genes were determined using real-time quantitative PCR method in 50 micromolar Epigallocatechin-3-gallate treated K-562 cells.

Results: Epigallocatechin-3-gallate treatment resulted in a significant decrease and increase the expression levels of autophagy-related genes and microRNAs targeting these genes in K-562 cells, respectively.

Conclusion: In the direction of the results, epigallocatechin-3-gallate was found to cause autophagy inhibition by genetic and epigenetic regulation in chronic myeloid leukemia cells. Considering the dual mechanism of autophagy, this result suggests that epigallocatechin-3-gallate is a chemical that is important for the nutrition of patients as well as its potential to be used as a drug.

Keywords

• Chronic Myeloid Leukemia,miRNA,Autophagy

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