Investigation of miR-21, miR-150, miR-155 Expression Levels in Chronic Myeloid Leukemia Patients

Abstract

Purpose: In this study, the relationship between CML and the expression levels of miR-21, miR-150, and miR-155, which could be used in the follow-up of CML patients, was investigated. Materials and Methods: RNA and miRNA were extracted from peripheral blood samples of patient and control samples. Targe-ted miRNA expression levels from cDNA samples were analyzed by real-time PCR method. Results: miRNA expression level was determined as 1.0 in the control group. In the newly diagnosed group, the mean miR-21 fold change was 0.6, miR-150 fold change was 0.3, and miR-155 fold change was 0.5. fold changes for miR-21, miR-150 and miR-155 were found to be 1.4, 0.5 and 2.4 fold, respectively, in the imatinib treatment group. In the nilotinib treatment group, miR-21 level was 3.5, miR-150 level 0.5, and miR-155 level 4.3. In the dasatinib treatment group, fold change was 0.8 for miR-21, 2.1 for miR-150, and 0.5 for miR-155. The mean miR-21 level was found to be 3.2, miR-150 level 1.0 and miR-155 level 2.8. MiR-150 levels were found to be lower in the newly diagnosed group, imatinib group and nilotinib group than in the control samples. This difference between the new diagnosis group (p=0.07484) and the nilotinib group (p=0.01541) is statistically signi-ficant. No significant difference was found between patients and controls in terms of miR-21 and miR-155 levels. Conclusion: These results support that miRNA-150 can be used as a parameter in the monitoring of treatment of CML patients and can contribute to the early detection of drug resistance.

Keywords

• chronic myeloid leukemia
• microRNA
• microRNA-21
• microRNA-150
• microRNA-155

Kronik Miyeloid Lösemi Hastaları’nda miR-21, miR-150, miR-155 Ekspresyon Düzeylerinin Araştırılması

Abstract

Amaç: MikroRNA (miRNA)’lar proteinlere translasyonu gerçekleşmeyen, hücrede biyolojik mekanizmaları etkileyen RNA molekülleridir. miRNA ekspresyon düzeylerinin qPCR ile belirlenmesi KML hastalarının tedavisinin takibinde bilgi verebilir. Bu çalışmada KML hastalarının takibinde kullanılabilecek miR-21, miR-150, miR-155 ekspresyonlarının KML ile ilişkisi araştırılması amaçlanmıştır. Araçlar ve Yöntem: Bu çalışma, imatinib, nilotinib, dasatinib ilaçlarıyla tedavi edilen 22 KML hastası ve 5 sağlıklı kontrolden elde edilen plazma örneklerinde yapıldı. Hasta ve kontrol periferik kan örneklerinden RNA ve miRNA eldesi Genaxxon miRNA pürifikasyon kiti kullanılarak gerçekleştirildi. Wiz ScriptcDNA Sentez Kiti kullanılarak cDNA eldesi yapıldı. cDNA örneklerinden hedeflenen miRNA ekspresyonlarının düzeyleri gerçek zamanlı PCR yöntemi ile analiz edildi. Bulgular: Kontrol grubu miRNA ekspresyon düzeyleri 1.0 olarak belirlendi. KML yeni tanı grubunda miR-21 kat değişimi ortalama 0.6, miR-150 kat değişimi 0.3 ve miR-155 kat değişimi 0.5 bulundu. İmatinib tedavi grubunda miR-21, miR-150 ve miR-155 için kat değişimi sırasıyla 1.4, 0.5 ve 2.4 kat olarak bulundu. Nilotinib tedavi grubunda miR-21 düzeyi 3.5, miR-150 düzeyi 0.5 ve miR-155 düzeyi 4.3 bulundu. Dasatinib tedavi grubunda miR-21 kat değişimi 0.8, miR-150 kat değişimi 2.1 ve miR-155 kat değişimi 0.5 bulundu. Yapılan çalışmada miR-21 düzeyi ortalama 3.2, miR-150 düzeyi 1.0 ve miR-155 düzeyi 2.8 bulundu. miR-150 düzeyleri yeni tanı grubunda, İmatinib grubunda ve Nilotinib grubunda kontrol örneklerinden daha düşük oranda bulundu. Bu fark istatistiksel olarak yeni tanı grubunda (p=0.07484) ve Nilotinib grubunda (p=0.01541) anlamlıdır. Hasta ve kontroller arasında miR-21 ve miR-155 düzeyleri yönünden istatiksel olarak anlamlı bir farklılık bulunmadı. Sonuç: Bu sonuçlar miRNA-150’nin KML hastalarının izleminde kullanılabileceğini ve ilaç direncinin erken dönemde tespitinde katkı sağlayabileceğini desteklemektedir.

Keywords

• kronik miyeloid lösemi
• mikroRNA
• mikroRNA-21
• mikroRNA-150
• mikroRNA-155

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