Megakaryoblast ve bazofil hücre tipine sahip Ph (+) KML hücre hatlarının Wnt ve Frizzled ilişkili gen ifadelerinin karşılaştırılması

Year 2021, Volume: 46 Issue: 1, 318 - 324, 31.03.2021

Melek Pehlivan , Hakkı Ogün Sercan

https://doi.org/10.17826/cumj.795833

Abstract

Amaç: Bu çalışmada blastik fazda farklı hücre serilerinde tutulum gösteren iki farklı Ph (+) kronik miyeloid lösemi (KML) hücre hattında, Wnt sinyal yolağı ana oyuncuları olan Wnt ve Frizzled genlerinin ifade farklılıklarının incelenmesi amaçlanmıştır.
Gereç ve Yöntem: Megakaryoblast (MEG-01) ve bazofil (KU812) hücre tipine sahip Ph (+) KML hücre hatları kültüre edildikten sonra, RNA izolasyonları gerçekleştirilmiştir. RNA konsantrasyonları spektrofotometrede belirlendikten sonra, cDNA çevrimi yapılmıştır. PCR ile Wnt ve Frizzled genlerinin ifade düzeyleri analiz edilmiş, iki hücre hattının Wnt ve Frizzled ekspresyon verileri birbirleri ile karşılaştırılmıştır.
Bulgular: Yapılan analizler sonrasında MEG-01 hücrelerinin Fzd1-9 genlerini, KU812’nin ise Fzd3, Fzd4, Fzd5, Fzd6, Fzd7, Fzd9 genlerini ifade ettiği gözlenmiştir. MEG-01 hücreleri Wnt2b ve Wnt3 genlerini eksprese ederken, KU812 hücrelerinin ise sadece Wnt3’ü eksprese ettiği saptanmıştır.
Sonuç: KU812 ve MEG-01 hücre hatlarının, Wnt sinyal genlerinin ifade düzeyleri açısından birbirinden farklılık gösterdiği belirlenmiştir. İki KML Ph (+) hücre hattında gözlenen Wnt sinyal genlerinin ifade düzeyleri arasındaki farklara, hücrenin orjini, füzyon bölgeleri ve hücre tipine özgü olabilecek sinyal ileti uyarı ve yolaklarındaki farklılıkların neden olabileceği düşünülmektedir. Bu nedenle model hücre hatları kullanılarak yapılan çalışmalar da birden fazla hücre hattının seçilmesi önerilmektedir.

Keywords

KML, Wnt Sinyal iletimi, Ph(+) hücre hatları

Supporting Institution

Dokuz Eylül Üniversitesi, İKÇÜ

Comparison of Wnt and Frizzled related gene expressions of Ph (+) CML cell lines with megakaryoblast and basophil cell types

Abstract

Purpose: The aim of this study was to investigate the expression changes in Wnt and Frizzled genes that initiate Wnt signaling in two different Ph (+) chronic myeloid leukemia (CML) cell lines.
Materials and Methods: Total RNA isolations were performed from Ph (+) CML cell lines with megakaryoblast and basophil cell types. RNA concentrations was evaluated by spectrophotometry and reverted to cDNA. Expression levels of Wnt and Frizzled genes were analyzed by PCR and data of two cell lines were compared with each other.
Results: In the analyzes, it was observed that MEG-01 cells expressed Fzd1-9 genes, while KU812 expressed Fzd3, Fzd4, Fzd5, Fzd6, Fzd7, Fzd9 genes. It was determined that MEG-01 cells expressed Wnt2b and Wnt3 genes, while KU812 cells expressed only Wnt3 gene.
Conclusion: KU812, MEG-01 cell lines differ from each other in terms of expression levels of Wnt signaling genes. It is thought that the differences between the expression levels of Wnt genes observed in the two CML Ph (+) cell lines may be caused by the differences in the cell's origin, fusion regions and cell type-specific signal transduction stimuli and pathways. Therefore, it is recommended to select more than one cell line in studies using model cell lines.

Keywords

CML, Wnt genes

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