Objective: Noncoding small RNAs play roles in physiological processes such as cell differentiation, growth, development, immune reactions, stress adaptation as well as complex diseases such as cancer and heart diseases. MIR223 is a specific miRNA to the hematopoietic system. Among the miRNAs associated with the pathogenesis of T-cell acute lymphoblastic leukemia (T-ALL), MIR223 was shown to have the highest expression level and functions as an oncomir. In this study we aimed to suppress MIR223 gene expression by using locked nucleic acid (LNA) in T-ALL cell lines as an alternative gene silencing technique and to show the silencing efficacy. Material and Methods: T-ALL cell lines (Jurkat and Molt4) were cultured and MIR223 specific LNA was applied in 100pmol and 150pmol concentrations for 24 hours and 48 hours. RNA was isolated from cells at both time points, followed by a stem loop polymerase chain reaction (PCR) for cDNA synthesis and the miRNA expression levels were evaluated by quantitative real time PCR. Results: In both cell lines a 150pmol application of LNA was compared to mock cells and the MIR223 expression was already suppressed at 24hours (73% in Jurkat (p=0,001) and 80%in Molt4 (p=0,04)). The suppression was continued at 48 hours in Molt4 cell line (p=0,005) where as there was no statistical significant difference in Jurkat at 48 hours. Conclusion: MIR223 has been identified as an oncomir and silencing the MIR223 with synthetic antisense oligos provide the opportunity to investigate the course and alternative therapy options of cancers with increased MIR223 expression, such as T-ALL. In this study LNA was used as an alternative RNA interference application in T-ALL cells and showed that LNA can effectively suppress gene expression when used in the short term.
|Destekleyen Kurum||Bu çalışma, İstanbul Üniversitesi Bilimsel Araştırma Projeleri Birimi tarafından desteklenmiştir.|
: 18 Mayıs 2020
|APA||Taşar, O , Sayi̇toğlu, M , Hatırnaz Ng, Ö . (2020). Hücre Serilerinde Kilitli Nükleik Asitler İle MIR223 Gen Sessizleştirmesi . Sağlık Bilimlerinde İleri Araştırmalar Dergisi , 3 (2) , 45-50 . Retrieved from https://dergipark.org.tr/tr/pub/sabiad/issue/55974/739221|