Investigation of the apoptotic and cell cycle effects of sorafenib and doxorubicin on URG4/URGCP in leukemia cells

Yavuz Dodurga; Levent Elmas; Mücahit Seçme; Nazlı Demirkıran; Çığır Biray Avcı; Gülseren Bağcı; Sevda Sağ; Lale Şatıroğlu Tufan

Makale detayları

Investigation of the apoptotic and cell cycle effects of sorafenib and doxorubicin on URG4/URGCP in leukemia cells

Yazarlar:

Yavuz Dodurga | Levent Elmas | Mücahit Seçme | Nazlı Demirkıran | Çığır Biray Avcı | Gülseren Bağcı | Sevda Sağ | Lale Şatıroğlu Tufan

Yayıncı:

Pamukkale University

Ana Başlıklar:

Biyokimya ve Hücre Biyolojisi (Diğer)

Dergi:

Pamukkale Tıp Dergisi Vol.17 Issue 3, pp.498-508

Belge Tipi:

Makale

Yayın Tarihi:

5/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001314161

pISSN:

1308-0865

Anahtar kelimeler:

Özet:

 Purpose: The aim of this study is to investigate the effects of anticancer drugs such as Sorafenib SOR and Doxorubicin DOX on URG4URGCP mRNA levels in K562 and HL-60 leukemia cells, elucidating their effects on apoptosis and cell cycle. The effects of these drugs on apoptosis and the cell cycle in leukemia cells have been explored. This research aims to understand the cellular effects of drugs used in leukemia treatment and contribute valuable insights to the drug development processes in leukemia therapy. Materials and methods: DOX and SOR were evaluated for their IC50 values in K562 and HL-60 cell lines using the CellTiter-Glo assay Promega, USA, based on ATP measurement. Total RNA isolation was performed using Trizol reagent in both control and dose groups of each treated cell line. Following RNA isolation, cDNAs were synthesized using the Transcriptor High Fidelity cDNA Synthesis Kit. Subsequently, changes in mRNA expression levels were examined using specific primers for URG4URGCP, Casp-3, Casp-8, Casp-9, FADD, DR4, TRADD, CCDN1, CDK4, CDK6, PTEN, P53, and Rel-A genes. Results: In the groups treated with Sorafenib, the IC50 dose for HL-60 cell line was calculated as 40 μM at the 24th hour, and for K562 cell line, it was calculated as 40 μM at the 48th hour. In the groups treated with Doxorubicin, the IC50 doses were calculated as 50 μM at the 48th hour for HL-60 cell line, and as 50 μM at the 72nd hour for K562 cell line. Significant increases were observed in the mRNA expression levels of Casp-8, Casp-9, TRADD, DR4, Rel A, and FADD genes in the groups treated with SOR, while a decrease was observed in the mRNA expression levels of URG4URGCP, CCDN1, CDK4, and CDK6 genes. In the groups treated with DOX, significant increases were observed in the fold changes of Casp-3, Casp-8, P53, and PTEN genes. However, a significant decrease in mRNA expression levels was observed in URG4URGCP, CCDN1, and CDK4 genes. Conclusion: As a result, it has been demonstrated that both SOR and DOX may play a role in regulating the mRNA expressions of URG4URGCP, Casp-3, Casp-8, Casp-9, CDK6, CDK4, CCND1, P53, PTEN, TRADD, DR4, Rel A, and FADD genes in HL-60 and K562 cells.

 Purpose: The aim of this study is to investigate the effects of anticancer drugs such as Sorafenib SOR and Doxorubicin DOX on URG4URGCP mRNA levels in K562 and HL-60 leukemia cells, elucidating their effects on apoptosis and cell cycle. The effects of these drugs on apoptosis and the cell cycle in leukemia cells have been explored. This research aims to understand the cellular effects of drugs used in leukemia treatment and contribute valuable insights to the drug development processes in leukemia therapy. Materials and methods: DOX and SOR were evaluated for their IC50 values in K562 and HL-60 cell lines using the CellTiter-Glo assay Promega, USA, based on ATP measurement. Total RNA isolation was performed using Trizol reagent in both control and dose groups of each treated cell line. Following RNA isolation, cDNAs were synthesized using the Transcriptor High Fidelity cDNA Synthesis Kit. Subsequently, changes in mRNA expression levels were examined using specific primers for URG4URGCP, Casp-3, Casp-8, Casp-9, FADD, DR4, TRADD, CCDN1, CDK4, CDK6, PTEN, P53, and Rel-A genes. Results: In the groups treated with Sorafenib, the IC50 dose for HL-60 cell line was calculated as 40 μM at the 24th hour, and for K562 cell line, it was calculated as 40 μM at the 48th hour. In the groups treated with Doxorubicin, the IC50 doses were calculated as 50 μM at the 48th hour for HL-60 cell line, and as 50 μM at the 72nd hour for K562 cell line. Significant increases were observed in the mRNA expression levels of Casp-8, Casp-9, TRADD, DR4, Rel A, and FADD genes in the groups treated with SOR, while a decrease was observed in the mRNA expression levels of URG4URGCP, CCDN1, CDK4, and CDK6 genes. In the groups treated with DOX, significant increases were observed in the fold changes of Casp-3, Casp-8, P53, and PTEN genes. However, a significant decrease in mRNA expression levels was observed in URG4URGCP, CCDN1, and CDK4 genes. Conclusion: As a result, it has been demonstrated that both SOR and DOX may play a role in regulating the mRNA expressions of URG4URGCP, Casp-3, Casp-8, Casp-9, CDK6, CDK4, CCND1, P53, PTEN, TRADD, DR4, Rel A, and FADD genes in HL-60 and K562 cells.

 Purpose: The aim of this study is to investigate the effects of anticancer drugs such as Sorafenib SOR and Doxorubicin DOX on URG4URGCP mRNA levels in K562 and HL-60 leukemia cells, elucidating their effects on apoptosis and cell cycle. The effects of these drugs on apoptosis and the cell cycle in leukemia cells have been explored. This research aims to understand the cellular effects of drugs used in leukemia treatment and contribute valuable insights to the drug development processes in leukemia therapy. Materials and methods: DOX and SOR were evaluated for their IC50 values in K562 and HL-60 cell lines using the CellTiter-Glo assay Promega, USA, based on ATP measurement. Total RNA isolation was performed using Trizol reagent in both control and dose groups of each treated cell line. Following RNA isolation, cDNAs were synthesized using the Transcriptor High Fidelity cDNA Synthesis Kit. Subsequently, changes in mRNA expression levels were examined using specific primers for URG4URGCP, Casp-3, Casp-8, Casp-9, FADD, DR4, TRADD, CCDN1, CDK4, CDK6, PTEN, P53, and Rel-A genes. Results: In the groups treated with Sorafenib, the IC50 dose for HL-60 cell line was calculated as 40 μM at the 24th hour, and for K562 cell line, it was calculated as 40 μM at the 48th hour. In the groups treated with Doxorubicin, the IC50 doses were calculated as 50 μM at the 48th hour for HL-60 cell line, and as 50 μM at the 72nd hour for K562 cell line. Significant increases were observed in the mRNA expression levels of Casp-8, Casp-9, TRADD, DR4, Rel A, and FADD genes in the groups treated with SOR, while a decrease was observed in the mRNA expression levels of URG4URGCP, CCDN1, CDK4, and CDK6 genes. In the groups treated with DOX, significant increases were observed in the fold changes of Casp-3, Casp-8, P53, and PTEN genes. However, a significant decrease in mRNA expression levels was observed in URG4URGCP, CCDN1, and CDK4 genes. Conclusion: As a result, it has been demonstrated that both SOR and DOX may play a role in regulating the mRNA expressions of URG4URGCP, Casp-3, Casp-8, Casp-9, CDK6, CDK4, CCND1, P53, PTEN, TRADD, DR4, Rel A, and FADD genes in HL-60 and K562 cells.

 Amaç: Çalışmanın amacı, Sorafenib SOR ve Doxorubicin DOX gibi antikanser ilaçlarının K562 ve HL-60 lösemi hücrelerinde URG4URGCP mRNA düzeyleri üzerindeki etkilerini inceleyerek, apoptoz ve hücre döngüsü üzerindeki etkilerini açıklığa kavuşturmaktır. Bu ilaçların lösemi hücrelerinde apoptoz ve hücre döngüsü üzerindeki etkileri araştırılmıştır. Bu araştırma, lösemi tedavisinde kullanılan ilaçların hücresel etkilerini anlamak ve lösemi tedavisinde ilaç geliştirme süreçlerine değerli bilgilerle katkıda bulunmayı amaçlamaktadır. Gereç ve yöntem: DOX ve SORin K562 ve HL-60 hücre hatlarında IC50 ATP ölçümüne dayanan CellTiter-Glo Promega, ABD tarafından değerlendirildi. Uygulama yapılan her iki hücre hattının kontrol ve doz gruplarında Trizol kimyasalı kullanılarak total RNA izolasyonu yapıldı. RNA izolasyonunun ardından Transcriptor High Fidelity cDNA Sentez Kiti ile cDNAlar sentezlendi. Daha sonra URG4URGCP, Casp-3, Casp-8, Casp-9, FADD, DR4, TRADD, CCDN1, CDK4, CDK6, PTEN, P53, Rel-A genlerine özgü primerler kullanılarak mRNA düzeyindeki ekspresyon değişiklikleri incelendi. Bulgular: Çalışmada Sorafenib uygulanan gruplarda IC50 dozu HL-60 hücre hattı için 24üncü saatte 40 μM, K562 hücre hattı için ise 48inci saatte 40 μM olarak hesaplandı. Doksorubisin uygulanan gruplarda ise IC50 dozları, HL-60 hücre hattı için 48inci saatte 50 μM, K562 hücre hattı için ise 72inci saatte 50 μM olarak hesaplanmıştır. SOR uygulanan gruplarda Casp-8, Casp-9, TRADD, DR4, Rel A ve FADD genlerin mRNA ekspresyon düzeylerindeki kat değişimlerinde önemli derecede artış gözlemlenirken, URG4URGCP, CCDN1, CDK4 ve CDK6 genlerinin mRNA ekspresyon düzeylerinde azalma olduğu gözlemlendi. DOX uygulanan gruplarda, Casp-3, Casp-8, P53 ve PTEN genlerinin kat değişimlerinde önemli derecede bir artış olduğu gözlemlenmiştir. Ancak, URG4URGCP, CCDN1 ve CDK4 genleri üzerinde mRNA ekspresyon düzeylerinde önemli bir azalma meydana gelmiştir. Sonuç: Sonuç olarak hem SOR hem de DOX in HL-60 ve K562 hücrelerinde URG4URGCP, Casp-3, Casp-8, Casp-9, CDK6, CDK4, CCND1, P53, PTEN, TRADD, DR4, Rel A ve FADD genlerin mRNA ekspresyonları düzenlenmesinde rol alabilecekleri gösterilmiştir.

Makale detayları

Investigation of the apoptotic and cell cycle effects of sorafenib and doxorubicin on URG4/URGCP in leukemia cells

Yazarlar:

Yavuz Dodurga | Levent Elmas | Mücahit Seçme | Nazlı Demirkıran | Çığır Biray Avcı | Gülseren Bağcı | Sevda Sağ | Lale Şatıroğlu Tufan

Yayıncı:

Pamukkale University

Ana Başlıklar:

Biyokimya ve Hücre Biyolojisi (Diğer)

Dergi:

Pamukkale Tıp Dergisi Vol.17 Issue 3, pp.498-508

Belge Tipi:

Makale

Yayın Tarihi:

5/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001314161

pISSN:

1308-0865

Doxorubicin

sorafenib

cell cycle

apoptosis

URG4/URGCPDoksorubisin

sorafenib

hücre döngüsü

apoptoz

URG4/URGCP

Sayı

Makale detayları

Yazarlar: Yavuz Dodurga | Levent Elmas | Mücahit Seçme | Nazlı Demirkıran | Çığır Biray Avcı | Gülseren Bağcı | Sevda Sağ | Lale Şatıroğlu Tufan

Yayıncı: Pamukkale University

Ana Başlıklar: Biyokimya ve Hücre Biyolojisi (Diğer)

Dergi: Pamukkale Tıp Dergisi Vol.17 Issue 3, pp.498-508

Belge Tipi: Makale

Yayın Tarihi: 5/2024

Belge Dili: İngilizce

Document ID (Org No.): 2001314161

pISSN: 1308-0865

Sayı

Yazarlar:

Yavuz Dodurga | Levent Elmas | Mücahit Seçme | Nazlı Demirkıran | Çığır Biray Avcı | Gülseren Bağcı | Sevda Sağ | Lale Şatıroğlu Tufan

Yayıncı:

Pamukkale University

Ana Başlıklar:

Biyokimya ve Hücre Biyolojisi (Diğer)

Dergi:

Pamukkale Tıp Dergisi Vol.17 Issue 3, pp.498-508

Belge Tipi:

Makale

Yayın Tarihi:

5/2024

Belge Dili:

İngilizce

Document ID (Org No.):

2001314161

pISSN:

1308-0865