Melek Pehlivan, Mustafa SOYÖZ, Hatice İlayhan KARAHAN ÇÖVEN, Burcu ÇERÇİ, Tülay KILIÇASLAN AYNA, Halil ATEŞ, Zeynep YÜCEL, Hakkı Ogün SERCAN

KML VE KML LÖSEMİK KÖK HÜCRESİ ARASINDA MikroRNA EKSPRESYON DEĞİŞİMLERİNİN DEĞERLENDİRİLMESİ

Amaç Kronik Miyeloid Lösemi (KML), hematopoetik kök hücreden (HKH) köken alan miyeloproliferatif bir has talıktır. MikroRNA' lar (miRNA), transkripsiyon sonrası gen ekspresyonunu düzenlenleyen küçük kodlama yan RNA’lardır. miRNA’lar KML’nin progresyonunda, lösemik kök hücre büyümesi ve tirozin kinaz inhibitörü (TKİ) direncinin gelişmesinde hücre homeostazisini etkilemektedirler. Bu çalışmada KML lösemik hücresi ve KML lösemik kök hücresi (LKH) arasında değişen miRNA ekspresyon profillerinin incelenmesi amaçlan mıştır. Gereç ve Yöntem KML hücre hattı olan K562 hücrelerinden, manye tik hücre ayrımlama (MACS) yöntemi kullanılarak CD34+CD38- lösemik kök hücreleri ayrımlanmıştır. Ayrımlanan LKH’lerin saflığının %85-92 arasında ol duğu akım sitometri yöntemi ile gösterilmiştir. K562 ve K562 LKH’leri arasında, gerçek zamanlı kantitatif PCR ile kanser kök hücre ilişkili 84 adet miRNA’nın ekspresyon değişimleri incelenmiştir. Bulgular K562 ve K562 LKH’leri arasında, kök hücre ilişkili ol duğu bilinen 84 adet miRNA’dan 7’sinin anlamlı dü zeyde değiştiğini gözledik (p

EVALUATION OF MicroRNA EXPRESSION CHANGES BETWEEN CML AND CML LEUKEMIC STEM CELL

Objective Chronic Myeloid Leukemia (CML) is a myeloprolifera tive disease, originating from the hematopoietic stem cells (HSC). MicroRNAs (miRNA) are small nonco ding RNAs that post transcriptionally regulate gene expression. miRNAs have been implicated in the progression of CML and may affect growth of leuke mic stem cell and cell homeostasis in the develop ment of tyrosine kinase inhibitors (TKI) resistance. In this study, we aimed to investigate miRNA expression profiles ranging from CML leukemic cell and CML leu kemic stem cell (LSC). Material and Methods CD34+CD38- leukemic stem cell population was iso lated from K562 cells by Magnetic Cell Separation (MACS) method. The purity of the separated LSCs were controlled by flow cytometry (85-92%). Expres sion changes of 84 cancer stem cell related miRNAs were analyzed by real-time quantitative PCR between K562 and K562 LCS. Results Data analysis revealed that 7 of 84 miRNAs were changed in K562 LKH as compared to K562 cells (p

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