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Role of EZH2 in regulating MET proto-oncogene in chronic lymphocytic leukemia
University of Skövde, School of Bioscience.
2018 (English)Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Mesenchymal–epithelial transition factor (c-MET) is a receptor tyrosine kinase (RTK) naturally activated by the binding of hepatocyte growth factor (HGF) and was shown to regulate many essential cellular processes including cell proliferation, motility, invasion, angiogenesis, and apoptosis. According to the recent ChIP sequencing data on EZH2 using CLL samples from the current lab, MET gene was shown to be direct target of EZH2 in CLL. In this study, we investigated if EZH2 is regulating the MET gene expression through histone methyl transferase activity (H3K27me3) in chronic lymphocytic leukemia (CLL).We performed down-regulated of EZH2 using siRNA and analyzed the expression levels of both EZH2 and MET gene at mRNA and protein levels using Real time-quantitative PCR, and western blot analysis respectively. In addition, we also analyzed if MET gene is regulated by DNA methylation using Pyrosequencing method. Our analysis showed that EZH2 directly regulated MET gene expression and DNA in the LINE1 region located at intron 2 regulates MET promoter, using CLL cell lines. Moreover, treatment of CLL cell with DNA methyl inhibitor drug, 5’-Deoxy 2’Azacytidine (DAC) drug induces the MET expression in CLL cell lines. Finally, this study shows that MET gene is regulated by both hypermethylation at intronic region and EZH2 binding on gene promoter resulting in gene silencing.

Place, publisher, year, edition, pages
2018.
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:his:diva-16140OAI: oai:DiVA.org:his-16140DiVA, id: diva2:1245074
Available from: 2018-09-06 Created: 2018-09-04 Last updated: 2018-09-06Bibliographically approved

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