The Tumor Suppressive mir-148a Is Epigenetically Inactivated in Classical Hodgkin Lymphoma

• Main Authors: Julia Paczkowska, Joanna Janiszewska, Julia Bein, Markus Schneider, Kinga Bednarek, Adam Ustaszewski, Sylvia Hartmann, Martin-Leo Hansmann, Maciej Giefing
• Format: Article
• Language: English
• Published: MDPI AG 2020-10-01
• Series: Cells
• Subjects: cHL; epigenetic; microRNA; DNA methylation; mir-148a
• Online Access: https://www.mdpi.com/2073-4409/9/10/2292

DNA methylation was shown previously to be a crucial mechanism responsible for transcriptional deregulation in the pathogenesis of classical Hodgkin lymphoma (cHL). To identify epigenetically inactivated miRNAs in cHL, we have analyzed the set of miRNAs downregulated in cHL cell lines using bisulfite pyrosequencing. We focused on miRNAs with promoter regions located within or <1000 bp from a CpG island. Most promising candidate miRNAs were further studied in primary Hodgkin and Reed-Sternberg (HRS) cells obtained by laser capture microdissection. Last, to evaluate the function of identified miRNAs, we performed a luciferase reporter assay to confirm miRNA: mRNA interactions and therefore established cHL cell lines with stable overexpression of selected miRNAs for proliferation tests. We found a significant reverse correlation between DNA methylation and expression levels of mir-339-3p, mir-148a-3p, mir-148a-5p and mir-193a-5 demonstrating epigenetic regulation of these miRNAs in cHL cell lines. Moreover, we demonstrated direct interaction between miR-148a-3p and <i>IL15</i> and <i>HOMER1</i> transcripts as well as between mir-148a-5p and <i>SUB1</i> and <i>SERPINH1</i> transcripts. Furthermore, mir-148a overexpression resulted in reduced cell proliferation in the KM-H2 cell line. In summary, we report that mir-148a is a novel tumor suppressor inactivated in cHL and that epigenetic silencing of miRNAs is a common phenomenon in cHL.