Premium
miR‐23a and miR‐23b Are Putative Regulators of KDM6A Protein Expression in Epithelial Mesenchymal Transition
Author(s) -
Kim Joelle,
Johnson Kelsey,
Taube Joseph
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.03704
Subject(s) - microrna , epithelial–mesenchymal transition , biology , messenger rna , demethylase , gene expression , cancer research , untranslated region , gene , downregulation and upregulation , genetics , histone
Cancer cell metastasis is the leading cause of cancer patient mortality. Tumor progression of epithelial cancers is facilitated by epithelial‐mesenchymal transition (EMT) and mesenchymal‐epithelial transition (MET), reverse processes that enable invasive cellular behavior. A key regulator of EMT/MET is lysine (K)‐specific demethylase 6A (KDM6A), an enzyme involved in epithelial gene expression that is abundant in epithelial cell phenotypes. We have previously determined that KDM6A is not dynamically regulated at the transcription level; we now aim to investigate whether post‐transcriptional mechanisms modulate KDM6A expression patterns. KDM6A mRNA transcripts are potentially regulated by specific miRNAs that inhibit expression of KDM6A protein, influencing EMT and downstream cellular morphology. To identify miRNAs that likely target KDM6A, eight databases were utilized to compile a list of ~160 potential miRNAs predicted to bind at the KDM6A 3′ untranslated region (UTR). Top miRNA candidates were then selected by number of deep sequencing reads, observed amounts in breast cancer cell lines, and frequency in patient data. miR‐23a, ‐23b, ‐145, and ‐199a were determined as potential regulators of KDM6A. The miRNAs were quantified by performing miRNA‐specific RT‐qPCR on RNA collected from TGFβ‐induced EMT and MET at various timepoints. Of the four miRNA candidates, miR‐23a and ‐23b were identified as likely post‐transcriptional regulators of KDM6A, as their relative quantification levels follow expected patterns opposing KDM6A expression throughout EMT/MET. Future directions aim to determine whether experimental manipulation of miR‐23a/b alters KDM6A protein expression and whether predicted binding sites at the KDM6A 3′ UTR are required. Cloning the gene for miR‐23a/b into a mammalian expression vector to observe miRNA‐overexpression effects on KDM6A expression can clarify KDM6A’s potential role in metastatic activity and can introduce new targets for research in clinical applications. Support or Funding Information Investigation of KDM6A function is supported through funding from Susan G. Komen and Jack G. and Norma Jean Folmar.