A Possible Role for WNT5A Hypermethylation in Pediatric Acute Lymphoblastic Leukemia

Turkish Journal of Haematology : Official Journal of Turkish Society of Haematology
Özden Hatırnaz NgMüge Sayitoğlu

Abstract

WNT5A is one of the most studied noncanonical WNT ligands and is shown to be deregulated in different tumor types. Our aim was to clarify whether hypermethylation might be the cause of low WNT5A mRNA levels and whether we could restore this downregulation by reversing the event. The expression of WNT5A mRNA was studied in a large acute lymphoblastic leukemia (ALL) patient group (n=86) by quantitative real-time PCR. The methylation status was detected by methylation-specific PCR (MSPCR) and bisulphate sequencing. In order to determine whether methylation has a direct effect on WNT5A expression, disease-representative cell lines were treated by 5'-aza-20-deoxycytidine. Here we designed a validation experiment of the WNT5A gene, which was previously examined and found to be differentially expressed by microarray study in 31 T-cell ALL patients. The expression levels were confirmed by quantitative real-time PCR and the expression levels were significantly lower in T-cell ALL patients than in control thymic subsets (p=0.007). MSPCR revealed that 86% of the patients were hypermethylated in the WNT5A promoter region. Jurkat and RPMI cell lines were treated with 5'-aza-20-deoxycytidine and WNT5A mRNA expression was restored after treat...Continue Reading

Citations

Mar 7, 2017·Turkish Journal of Medical Sciences·Sinem FirtinaMüge Sayitoğlu
Jun 24, 2020·Journal of Leukocyte Biology·Sheila C Soares-LimaFlávia R G Carneiro
Jul 7, 2018·Journal of Cell Communication and Signaling·Sukalpa ChattopadhyaySujata Law

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Methods Mentioned

BETA
FACS
density-gradient centrifugation
PCR
electrophoresis

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CLC Main Workbench
GraphPad Prism
SPSS

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