TR-Dizin İndeksli Yayınlar Koleksiyonu

Permanent URI for this collectionhttps://hdl.handle.net/20.500.12573/396

Browse

Filters

20223

Settings

Author: search.filters.author.Okur, TuğbaPublication Index: search.filters.publicationIndex.TR-Dizin

Search Results

Now showing 1 - 3 of 3
  • Research Project
    Investigation of the Antitumor Effect of Targeting PI3K-AKT-mTOR Pathway and Histone Deacetylase Enzymes on Acute Myeloid Leukemia Cells
    (2022) Şansaçar, Merve; Akçok, Emel Başak Gencer; Okur, Tuğba; Karaca, Münevver
    Acute Myeloid Leukemia (AML) is a disease characterized by the accumulation of immature myeloid cells called blasts in the peripheral blood, bone marrow, spleen, and liver, eventually leading to hematopoietic malignancy. In addition to genetic abnormalities, important cellular pathways such as PI3K/AKT/mTOR, Wnt, Notch, STAT3, Hedgehog have been reported to play a role in the pathogenesis of AML. Histone deacetylase (HDAC) inhibitors have promising anticancer activity for AML. In this study, it was aimed to investigate the effect of inhibition of the PI3K/AKT/mTOR pathway and HDAC inhibition on the molecular mechanism underlying this disease using cell lines of different AML subgroups, MOLM-13 and CMK cell lines. For this purpose, the effects of PI3K inhibitor LY294002 and HDAC inhibitors (SAHA, PCI-3501 and Tubastatin A) and their combinations were investigated. Cell proliferation was determined by MTT cell cytotoxicity test and apoptosis rates were determined by Annexin-V/PI double staining method, and the effects of drugs on the cell cycle were determined by PI staining. The level of LC3B protein, a marker of autophagy, was confirmed at the molecular level by western blot. The inhibitors used decreased cell viability at low micromolar concentrations on both cell lines. It was determined that the LY294002+SAHA combination treatment showed a 50% reduction in cell proliferation in MOLM-13 cells and a 25% decrease in CMK cells. LY294002+Tubastatin A treatment has been shown to reduce cell proliferation in MOLM-13 and CMK cells by 65% and 40%, respectively. Our results showed that combinations of LY294002 and HDAC inhibitor resulted in G0/G1 phase arrest in MOLM-13 cells compared to control cells. On the other hand, CMK cells treated with combinations of LY294002+SAHA, LY294002+PCI-3501 and LY294002+Tubastatin A were arrested in G2/M, G2/M and G0/G1 phase, respectively. The effect of the combinations on apoptotic cell death was investigated. LC3B protein expression level was checked as a result of combination therapy. Considering the effects of HDAC enzymes on both AML and different cancers, HDAC inhibition is an important and high-potential target for AML. Therefore, investigation of the PI3K/AKT/mTOR pathway and inhibition of HDACs in different subgroups may provide insight into the mechanisms leading to the pathogenesis of AML. Consequently, it is hoped that this inhibition of PI3K/AKT/mTOR and HDAC will lead to a more specific combination of targeted therapy that results in the abolition of AML.
  • Research Project
    PI3K-AKT-mTOR Yolağı ve Histon Deasetilaz Enzimlerinin Hedeflenmesinin Akut Myeloid Lösemi Hücreleri Üzerine Antitümör Etkisinin İncelenmesi
    (2022) Şansaçar, Merve; Akçok, Emel Başak Gencer; Okur, Tuğba; Karaca, Münevver
    Acute Myeloid Leukemia (AML) is a disease characterized by the accumulation of immature myeloid cells called blasts in the peripheral blood, bone marrow, spleen, and liver, eventually leading to hematopoietic malignancy. In addition to genetic abnormalities, important cellular pathways such as PI3K/AKT/mTOR, Wnt, Notch, STAT3, Hedgehog have been reported to play a role in the pathogenesis of AML. Histone deacetylase (HDAC) inhibitors have promising anticancer activity for AML. In this study, it was aimed to investigate the effect of inhibition of the PI3K/AKT/mTOR pathway and HDAC inhibition on the molecular mechanism underlying this disease using cell lines of different AML subgroups, MOLM-13 and CMK cell lines. For this purpose, the effects of PI3K inhibitor LY294002 and HDAC inhibitors (SAHA, PCI-3501 and Tubastatin A) and their combinations were investigated. Cell proliferation was determined by MTT cell cytotoxicity test and apoptosis rates were determined by Annexin-V/PI double staining method, and the effects of drugs on the cell cycle were determined by PI staining. The level of LC3B protein, a marker of autophagy, was confirmed at the molecular level by western blot. The inhibitors used decreased cell viability at low micromolar concentrations on both cell lines. It was determined that the LY294002+SAHA combination treatment showed a 50% reduction in cell proliferation in MOLM-13 cells and a 25% decrease in CMK cells. LY294002+Tubastatin A treatment has been shown to reduce cell proliferation in MOLM-13 and CMK cells by 65% and 40%, respectively. Our results showed that combinations of LY294002 and HDAC inhibitor resulted in G0/G1 phase arrest in MOLM-13 cells compared to control cells. On the other hand, CMK cells treated with combinations of LY294002+SAHA, LY294002+PCI-3501 and LY294002+Tubastatin A were arrested in G2/M, G2/M and G0/G1 phase, respectively. The effect of the combinations on apoptotic cell death was investigated. LC3B protein expression level was checked as a result of combination therapy. Considering the effects of HDAC enzymes on both AML and different cancers, HDAC inhibition is an important and high-potential target for AML. Therefore, investigation of the PI3K/AKT/mTOR pathway and inhibition of HDACs in different subgroups may provide insight into the mechanisms leading to the pathogenesis of AML. Consequently, it is hoped that this inhibition of PI3K/AKT/mTOR and HDAC will lead to a more specific combination of targeted therapy that results in the abolition of AML.
  • Research Project
    Pi3k-Akt-Mtor Yolağı Ve Histon Deasetilaz Enzimlerinin Hedeflenmesinin Akut Myeloid Lösemi Hücreleri Üzerine Antitümör Etkisinin Incelenmesi
    (TÜBİTAK, 2022) Gencer Akçok, Emel Başak; Şansaçar, Merve; Karaca, Münevver; Okur, Tuğba
    Akut Miyeloid Lösemi (AML), periferik kan, kemik iliği, dalak ve karaciğerde blast adı verilen_x000D_ olgunlaşmamış miyeloid hücrelerin birikmesiyle karakterize edilen ve sonunda hematopoietik_x000D_ maligniteye yol açan bir hastalıktır. Genetik anormalliklerin yanı sıra PI3K/AKT/mTOR, Wnt,_x000D_ Notch, STAT3, Hedgehog gibi önemli hücresel yolakların AML patogenezinde rol oynadığı_x000D_ bildirilmiştir. Histon deasetilaz (HDAC) inhibitörleri, AML için umut verici antikanser aktiviteye_x000D_ sahiptir. Çalışmada, PI3K/AKT/mTOR yolunun inhibisyonunun ve HDAC inhibisyonunun farklı_x000D_ AML alt gruplarının hücre hatları olan MOLM-13 ve CMK hücre hatları kullanılarak bu_x000D_ hastalığın altında yatan moleküler mekanizma üzerindeki etkisinin araştırılması amaçlamıştır._x000D_ Bu amaçla PI3K inhibitörü LY294002 ve HDAC inhibitörleri (SAHA, PCI-3501 ve Tubastatin A)_x000D_ ve bunların kombinasyonlarının etkisi incelenmiştir. Hücre proliferasyonu MTT hücre_x000D_ sitotoksisite testi ile apoptoz oranları ise Annexin-V/PI çift boyama yöntemi ile belirlenmiş,_x000D_ ilaçların hücre döngüsüne olan etkileri de PI boyaması ile belirlenmiştir. Otofaji belirteci olan_x000D_ LC3B protein seviyesi moleküler düzeyde western blot ile doğrulanmıştır._x000D_ Kullanılan inhibitörler her iki hücre hattı üzerinde düşük mikromolar konsantrasyonda hücre_x000D_ canlılığını azaltmıştır. Sonuçlar LY294002+SAHA kombinasyon tedavisinin MOLM-13_x000D_ hücrelerinde hücre proliferasyonunu %50, CMK hücrelerinde ise %25 azalma gösterdiği_x000D_ belirlenmiştir. LY294002+Tubastatin A tedavisi, MOLM-13 ve CMK hücrelerinde hücre_x000D_ proliferasyonunu sırasıyla %65 ve %40 oranında azalttığını göstermiştir. Sonuçlarımız,_x000D_ LY294002 ve HDAC inhibitör kombinasyonlarının kontrol hücrelerine kıyasla MOLM-13_x000D_ hücrelerinde G1 fazı tutuklanmasıyla sonuçlandığını gösterdi. Öte yandan, LY294002+SAHA,_x000D_ LY294002+PCI-3501 ve LY294002+Tubastatin A kombinasyonları ile tedavi edilen CMK_x000D_ hücreleri, sırasıyla G2/M, G2/M ve G1 fazında tutuklanmıştır. Kombinasyonların apoptotik_x000D_ hücre ölümü üzerine etkisine bakılmış, LC3BII/I protein ifade düzeyi kombinasyon tedavisi_x000D_ sonucunda incelenmiştir._x000D_ HDAC enzimlerinin hem AML hem de farklı kanserler üzerindeki etkileri düşünüldüğünde,_x000D_ HDAC inhibisyonu AML için önemli ve yüksek potansiyelli bir hedeftir. Bu nedenle_x000D_ PI3K/AKT/mTOR yolağı ve HDAC'lerin farklı alt gruplarda inhibisyonunun araştırılması,_x000D_ AML'nin patogenezine yol açan mekanizmalar hakkında fikir verebilir. Sonuç olarak,_x000D_ PI3K/AKT/mTOR ve HDAC'nin bu inhibisyonunun, AML'nin ortadan kaldırılmasıyla_x000D_ sonuçlanan daha spesifik bir kombinasyon hedefli tedaviye yol açacağı umulmaktadır.