AN INVESTIGATION ON THE EFFECTS OF SIRT5 MODULATORS ON SIRT5 AND CYTOCHROME C PROTEIN EXPRESSIONS IN K562 CHRONIC MYELOID LEUKEMIA CELL LINE

Abstract

Objective: SIRT5 is a mitochondrial protein that removes acetyl, malonyl and succinyl groups from lysine moieties in target proteins and interacts with cytochrome c and causes its deacetylation. There is no study on the effects of SIRT5 in K562 chronic myeloid leukemia cells. Resveratrol and Suramin are known to play a role in modulating the deacetylase and desuccinylase activities of SIRT5. It has been reported that Resveratrol induces apoptosis of K562 cells but effects of Suramin on the apoptosis of K562 cells are largely unknown. In this study, it was aimed to elucidate the effects of SIRT5 modulators Resveratrol and Suramin on proliferation and apoptosis of K562 cells and on SIRT5 and cytochrome c protein, a known target of SIRT5.
Material and Method: K562 chronic myeloid leukemia cells were treated with increasing concentrations of suramin and resveratrol, cell proliferation was determined by MTT assay and BrdU incorporation. Apoptosis was determined with Annexin V staining by Flow cytometry. Western Blot analysis was performed to determine the effect of resveratrol and suramin on SIRT5 and Cytochrome c protein expression levels.
Result and Discussion: Our results showed that suramin did not affect SIRT5 and cytochrome c protein expressions significantly and resveratrol decreased SIRT5 and increased cytochrome c expression. Suramin did not cause any changes on the apoptosis of K562 cells. Resveratrol decreased cell proliferation and induced apoptosis of K562 cells in accordance with the literature. The SIRT5-lowering effect of Resveratrol may have mediated its apoptotic effects.

Keywords

• Apoptosis
• K562
• Resveratrol
• SIRT5
• Suramin

K562 KRONİK MİYELOİD LÖSEMİ HÜCRE HATTINDA SIRT5 MODÜLATÖRLERİNİN SIRT5 VE SİTOKROM C PROTEİN EKSPRESYONLARI ÜZERİNE ETKİLERİNİN ARAŞTIRILMASI

Abstract

Amaç: SIRT5, hedef proteinlerdeki lizin rezidülerinden, asetil, malonil ve süksinil gruplarını uzaklaştıran ve sitokrom c ile etkileşerek, onun deasetilasyonuna neden olan bir mitokondriyal proteindir. SIRT5'in K562 kronik miyeloid lösemi hücrelerindeki etkilerine ilişkin bir çalışma bulunmamaktadır. Resveratrol ve Suramin'in SIRT5'in deasetilaz ve desüksinilaz aktivitelerini modüle etmede rol oynadığı bilinmektedir. Resveratrol'ün K562 hücrelerinin apoptozunu indüklediği bildirilmiştir. Ancak Suramin'in K562 hücrelerinin apoptozu üzerindeki etkileri büyük ölçüde bilinmemektedir. Bu çalışmada, SIRT5 modülatörleri Resveratrol ve Suramin'in K562 hücrelerinin proliferasyonu ve apoptozu ile SIRT5 ve SIRT5'in bilinen bir hedefi olan sitokrom c proteini üzerindeki etkilerinin aydınlatılması amaçlanmıştır.
Gereç ve Yöntem: K562 kronik miyeloid lösemi hücrelerine artan konsantrasyonlarda Suramin ve Resveratrol uygulandı. Hücre proliferasyonu MTT analizi ve BrdU inkoporasyon yöntemi ile belirlendi. Apoptoz, Akım sitometrisi ile Annexin V boyaması ile belirlendi. Resveratrol ve Suramin'in SIRT5 ve Sitokrom c protein ekspresyon seviyeleri üzerindeki etkisini belirlemek için Western Blot analizi yapıldı.
Sonuç ve Tartışma: Sonuçlarımız, Suramin'in SIRT5 ve sitokrom c protein ekspresyonlarını önemli ölçüde etkilemediğini ve Resveratrol'ün SIRT5'i azalttığını ve sitokrom c ekspresyonunu artırdığını göstermiştir. Suramin, K562 hücrelerinin apoptozunda herhangi bir değişikliğe neden olmamıştır. Resveratrol, literatüre uygun olarak hücre proliferasyonunu azaltmış ve K562 hücrelerinin apoptozunu indüklemiştir. Resveratrolün, SIRT5 protein ekspresyonunu azaltıcı etkisi ile apoptotik etkilerine aracılık etmiş olabileceği düşünülmektedir

Keywords

• Apoptoz
• K562
• Resveratrol
• SIRT5
• Suramin

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