Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia

ENGÜR, Selin; Koca, Duygu; Kiraz, Yağmur; ULU, Gizem Tuğçe; Baran, Yusuf; ÇEKDEMIR, DEMET; Haştar, Nurcan

doi:10.4274/tjh.galenos.2019.2019.0083

Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia

Duygu KOCA, (İzmir Yüksek Teknoloji Enstitüsü, Moleküler Biyoloji ve Genetik Anabilim Dalı, İzmir, Türkiye)

Nurcan HASTAR, (İzmir Yüksek Teknoloji Enstitüsü, Moleküler Biyoloji ve Genetik Anabilim Dalı, İzmir, Türkiye)

Selin ENGÜR, (Anadolu Üniversitesi, Eczacılık Fakültesi, Farmakoloji Anabilim Dalı, Eskişehir, Türkiye)

Yağmur KİRAZ, (İzmir Yüksek Teknoloji Enstitüsü, Moleküler Biyoloji ve Genetik Anabilim Dalı, İzmir, Türkiye)

Gizem Tuğçe ULU, (İzmir Yüksek Teknoloji Enstitüsü, Moleküler Biyoloji ve Genetik Anabilim Dalı, İzmir, Türkiye)

Demet ÇEKDEMİR, (Sakarya Üniversitesi Tıp Fakültesi, Hematoloji Anabilim Dalı, Sakarya, Türkiye)

Yusuf BARAN (İzmir Yüksek Teknoloji Enstitüsü, Moleküler Biyoloji ve Genetik Anabilim Dalı, İzmir, Türkiye)

Turkish Journal of Hematology

2 0

Yıl: 2020 Cilt: 37 Sayı: 1 Sayfa Aralığı: 5 - 12 Metin Dili: İngilizce DOI: 10.4274/tjh.galenos.2019.2019.0083 İndeks Tarihi: 16-10-2020

Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia

Öz:

Objective: Acute myeloid leukemia (AML) is a complex disease affected by both genetic and epigenetic factors. Histone methylation and demethylation are types of epigenetic modification in chromatin remodeling and gene expression. Abnormal expression of histone demethylases is indicated in many types of cancer including AML. Although many commercial drugs are available to treat AML, an absolute cure has not been discovered yet. However, inhibition of demethylases could be a potential cure for AML. Methylstat is a chemical agent that inhibits the Jumonji C domain-containing demethylases. Materials and Methods: The cytotoxic and apoptotic effects of methylstat and doxorubicin on HL-60 cells were detected by MTT cell viability assay, double staining of treated cells with annexin-V/ propidium iodide, and caspase-3 activity assay. Mitochondrial activity was analyzed using JC-1 dye. The expression levels of the BCL2 and BCL2L1 anti-apoptotic genes in HL-60 cells were determined using real-time polymerase chain reaction (PCR). Lastly, the cytostatic effect was determined by cell cycle analysis. Results: In our research, cytotoxic, cytostatic, and apoptotic effects of methylstat on human HL-60 cells were investigated. Cytotoxic and cytostatic analyses revealed that methylstat decreased cell proliferation in a dose-dependent cytotoxic manner and arrested HL60 cells in the G2/M and S phases. Methylstat also induced apoptosis through the loss of mitochondrial membrane potential and increases in caspase-3 enzyme activity. The expression levels of BCL2 and BCL2L1 were also decreased according to real-time PCR results. Finally, the combination of methylstat with doxorubicin resulted in synergistic cytotoxic effects on HL-60 cells. Conclusion: Taken together, these results demonstrate that methylstat may be a powerful candidate as a drug component of AML treatment protocols.

Anahtar Kelime:

Akut Myeloid Lösemide Demetilaz İçeren Jumanji C Domainin İnhibisyonunun Terapötik Potansiyeli

Öz:

Amaç: Akut myeloid lösemi (AML) hem genetik hem de epigenetik faktörlerden etkilenen kompleks bir hastalıktır. Histon metilasyonu ve demetilasyonu, kromatin yeniden yapılanması ve gen ekspresyonu tayinindeki önemli epigenetik modifikasyonlarındandır. Histon demetilazların anormal ekspresyonu, AML dahil birçok kanser türünde etkilidir. AML’yi tedavi etmek için birçok ticari ilaç mevcut olmasına rağmen, kesin tedavisi bulunmamaktadır. Bu nedenle, demetilazların inhibisyonu AML için potansiyel bir tedavi olabilir. Methylstat, demetilaz içeren Jumonji C domain-inhibe eden bir kimyasal maddedir.Gereç ve Yöntemler: Methylstat ve doksorubisinin HL-60 hücreleri üzerindeki sitotoksik ve apoptotik etkisi, sırasıyla MTT canlılık testi, annexin-V/propidium iyodür çift boyaması ve kaspaz-3 aktivite testi ile saptandı. Ayrıca, hücrelerdeki mitokondriyal aktivite JC-1 boyası kullanılarak analiz edildi. HL-60 hücrelerinde BCL2 ve BCL2L1 apoptotik olmayan genlerin ekspresyon seviyeleri, gerçek zamanlı polimeraz zincir reaksiyonu (PZR) kullanılarak belirlendi. Son olarak, sitostatik etki hücre döngüsü analizi ile belirlenmiştir.Bulgular: Araştırmamızda, methylstatın HL-60 hücreleri üzerindeki sitotoksik, sitostatik ve apoptotik etkileri olduğu belirlenmiştir. Sitotoksik ve sitostatik analizlerde methylstatın, HL-60 hücrelerinde doza bağlı hücre çoğalmasını azalttığını ve G2/M ve S fazlarındaki sitotoksik etkisinin arttığı ortaya koydu. Methylstat ayrıca, mitokondriyal membran potansiyelini düşürdüğü ve kaspaz-3 enzim aktivitesindeki artışlarla apoptozu indüklemiştir. Ayrıca, gerçek zamanlı PZR sonuçlarına göre BCL2 ve BCL2L1 genlerinin ekspresyon seviyeleri azalmaktadır. Son olarak, methylstatın doksorubisin ile kombinasyonu, HL-60 hücreleri üzerinde sinerjik sitotoksik etkilere yol açmıştır.Sonuç: Methylstatın AML için tam bir tedavi bulmak için bir ilaç bileşeni olarak güçlü bir aday olabileceğini göstermiştir.

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	Koca D, Haştar N, ENGÜR S, Kiraz Y, ULU G, ÇEKDEMIR D, Baran Y (2020). Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. , 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
Chicago	Koca Duygu,Haştar Nurcan,ENGÜR Selin,Kiraz Yağmur,ULU Gizem Tuğçe,ÇEKDEMIR DEMET,Baran Yusuf Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. (2020): 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
MLA	Koca Duygu,Haştar Nurcan,ENGÜR Selin,Kiraz Yağmur,ULU Gizem Tuğçe,ÇEKDEMIR DEMET,Baran Yusuf Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. , 2020, ss.5 - 12. 10.4274/tjh.galenos.2019.2019.0083
AMA	Koca D,Haştar N,ENGÜR S,Kiraz Y,ULU G,ÇEKDEMIR D,Baran Y Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. . 2020; 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
Vancouver	Koca D,Haştar N,ENGÜR S,Kiraz Y,ULU G,ÇEKDEMIR D,Baran Y Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. . 2020; 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
IEEE	Koca D,Haştar N,ENGÜR S,Kiraz Y,ULU G,ÇEKDEMIR D,Baran Y "Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia." , ss.5 - 12, 2020. 10.4274/tjh.galenos.2019.2019.0083
ISNAD	Koca, Duygu vd. "Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia". (2020), 5-12. https://doi.org/10.4274/tjh.galenos.2019.2019.0083

APA	Koca D, Haştar N, ENGÜR S, Kiraz Y, ULU G, ÇEKDEMIR D, Baran Y (2020). Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. Turkish Journal of Hematology, 37(1), 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
Chicago	Koca Duygu,Haştar Nurcan,ENGÜR Selin,Kiraz Yağmur,ULU Gizem Tuğçe,ÇEKDEMIR DEMET,Baran Yusuf Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. Turkish Journal of Hematology 37, no.1 (2020): 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
MLA	Koca Duygu,Haştar Nurcan,ENGÜR Selin,Kiraz Yağmur,ULU Gizem Tuğçe,ÇEKDEMIR DEMET,Baran Yusuf Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. Turkish Journal of Hematology, vol.37, no.1, 2020, ss.5 - 12. 10.4274/tjh.galenos.2019.2019.0083
AMA	Koca D,Haştar N,ENGÜR S,Kiraz Y,ULU G,ÇEKDEMIR D,Baran Y Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. Turkish Journal of Hematology. 2020; 37(1): 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
Vancouver	Koca D,Haştar N,ENGÜR S,Kiraz Y,ULU G,ÇEKDEMIR D,Baran Y Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia. Turkish Journal of Hematology. 2020; 37(1): 5 - 12. 10.4274/tjh.galenos.2019.2019.0083
IEEE	Koca D,Haştar N,ENGÜR S,Kiraz Y,ULU G,ÇEKDEMIR D,Baran Y "Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia." Turkish Journal of Hematology, 37, ss.5 - 12, 2020. 10.4274/tjh.galenos.2019.2019.0083
ISNAD	Koca, Duygu vd. "Therapeutic Potentials of Inhibition of Jumonji C Domaincontaining Demethylases in Acute Myeloid Leukemia". Turkish Journal of Hematology 37/1 (2020), 5-12. https://doi.org/10.4274/tjh.galenos.2019.2019.0083