Knockdown of Stromal Interaction Molecule 1 (STIM1) Suppresses Acute Myeloblastic Leukemia-M5 Cell Line Survival Through Inhibition of Reactive Oxygen Species ActivitiesEman Salem Algariri1, Rabiatul Basria S. M. N. Mydin2, Emmanuel Jairaj Moses2, Simon Imakwu Okekpa3, Nur Arzuar Abdul Rahim4, Narazah Mohd Yusoff41Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Biomedical Science, Pulau Pinang, Malaysia; Hadhramout University, Faculty of Medicine and Health Sciences, Department of Basic Medical Sciences, Hadhramout, Yemen
2Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Biomedical Science, Pulau Pinang, Malaysia
3Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Biomedical Science, Pulau Pinang, Malaysia; Ebonyi State University, Faculty of Health Sciences, Department of Medical Laboratory Science, Ebonyi, Nigeria
4Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Clinical Medicine, Pulau Pinang, Malaysia
Objective: This study aimed to investigate the role of the stromal interaction molecule 1 (STIM1) gene in the survival of the acute myeloblastic leukemia (AML)-M5 cell line (THP-1).
Materials and Methods: The STIM1 effect was assessed via dicersubstrate siRNA-mediated STIM1 knockdown. The effect of STIM1 knockdown on the expression of AKT and MAPK pathway-related genes and reactive oxygen species (ROS) generation-related genes was tested using real-time polymerase chain reaction. Cellular functions, including ROS generation, cell proliferation, and colony formation, were also evaluated following STIM1 knockdown.
Results: The findings revealed that STIM1 knockdown reduced intracellular ROS levels via downregulation of NOX2 and PKC. These findings were associated with the downregulation of AKT, KRAS, MAPK, and CMYC. BCL2 was also downregulated, while BAX was upregulated following STIM1 knockdown. Furthermore, STIM1 knockdown reduced THP-1 cell proliferation and colony formation.
Conclusion: This study has demonstrated the role of STIM1 in promoting AML cell proliferation and survival through enhanced ROS generation and regulation of AKT/MAPK-related pathways. These findings may help establish STIM1 as a potential therapeutic target for AML treatment.
Keywords: STIM1, Reactive oxygen species, Survival, Proliferation, Therapeutic targets in acute myeloid leukemia
Stromal Etkileşim Molekülü 1’in (STIM1) Bozulması Reaktif Oksijen Türevleri Aktivitesinin İnhibisyonu Aracılığı ile Akut Myeloblastik Lösemi-M5 Hücre Dizilerinin Sağkalımını BaskılarEman Salem Algariri1, Rabiatul Basria S. M. N. Mydin2, Emmanuel Jairaj Moses2, Simon Imakwu Okekpa3, Nur Arzuar Abdul Rahim4, Narazah Mohd Yusoff41Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Biomedical Science, Pulau Pinang, Malaysia; Hadhramout University, Faculty of Medicine and Health Sciences, Department of Basic Medical Sciences, Hadhramout, Yemen
2Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Biomedical Science, Pulau Pinang, Malaysia
3Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Biomedical Science, Pulau Pinang, Malaysia; Ebonyi State University, Faculty of Health Sciences, Department of Medical Laboratory Science, Ebonyi, Nigeria
4Universiti Sains Malaysia, Advanced Medical and Dental Institute, Department of Clinical Medicine, Pulau Pinang, Malaysia
Amaç: Bu çalışmada stromal etkileşim molekülü 1 (STIM1) geninin akut myeloblastik lösemi (AML)-M5 hücre dizisinin (THP-1) sağkalımındaki rolü araştırıldı.
Gereç ve Yöntem: STIM1 etkisi, yarıcı substrat siRNA aracılı STIM1 bozulması yoluyla değerlendirildi. STIM1 bozulmasının AKT ve MAPK yolakları ve reaktif oksijen türevleri (ROS) üretimi ile ilişkili genlerin ekspresyonu üzerindeki etkisi RT-qPCR ile test edildi. Hücresel fonksiyonlar, ROS üretimi, hücre proliferasyonu ve koloni oluşumu dahil, STIM1 bozulmasını takiben değerlendirildi.
Bulgular: Sonuçlar STIM1 bozulmasının, NOX2 ve PKC aşağı düzenlemesi ile hücre içi ROS seviyesini düşürdüğünü ortaya koydu. Bu bulgular AKT, KRAS, MAPK aşağı düzenlemesi ile birlikteydi ve ayrıca CMYC, BCL2’de aşağı düzenlenmişti, BAX ise STIM1 bozulmasını takiben yukarı düzenlenmişti. Ek olarak STIM1 bozulması THP-1 hücre proliferasyonu ve koloni oluşumunu da azaltmıştı.
Sonuç: Bu çalışma, STIM1'in artmış ROS üretimi ve AKT/MAPK ile ilişkili yolların düzenlenmesi yoluyla AML hücre proliferasyonunu ve sağkalımını desteklemedeki rolünü göstermiştir. Bu bulgular, STIM1'in AML tedavisi için potansiyel bir terapötik hedef oluşturulmasına yardımcı olabilir.
Anahtar Kelimeler: STIM1, Reaktif oksijen türevleri, Sağkalım, Proliferasyon, Akut Myeloid lösemide terapötik hedefler
Eman Salem Algariri, Rabiatul Basria S. M. N. Mydin, Emmanuel Jairaj Moses, Simon Imakwu Okekpa, Nur Arzuar Abdul Rahim, Narazah Mohd Yusoff. Knockdown of Stromal Interaction Molecule 1 (STIM1) Suppresses Acute Myeloblastic Leukemia-M5 Cell Line Survival Through Inhibition of Reactive Oxygen Species Activities. Turk J Hematol. 2023; 40(1): 11-17
Corresponding Author: Rabiatul Basria S. M. N. Mydin, Malaysia |
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