IL32 overexpression is driven by DNA hypomethylation and contributes to an extracellular matrix (ECM) remodeling phenotype in EpCAM-/CD49f-enriched breast cancer cells

Elayne M. Benson, Megan A. Wilson, Emma V. Gray, Paris L. Rizzo, Maria Ouzounova, Hasan Korkaya, Austin Y. Shull

Research output: Contribution to journalArticlepeer-review

Abstract

Basal-like breast cancers typically correspond with increased enrichment of EpCAM-/CD49f- cancer stem cells (CSC) and a propensity toward metastasis. However, the molecular mechanisms underlying these general characteristics are not well understood. To provide further insight concerning CSCs and their intrinsic metastatic mechanisms, we compared the 450K DNA methylation profile of EpCAM-/CD49f- poor breast cancer cell lines to that of EpCAM-/CD49f- enriched breast cancer cell lines. From our results, we were able to determine and highlight IL32 as a gene whose promoter is hypomethylated in EpCAM-/CD49f- enriched cell lines. The hypomethylated IL32 promoter corresponded with increased IL32 expression in both cell lines and basal-like breast cancer patients from The Cancer Genome Atlas (TCGA) database. Interestingly, increased IL32 expression preferentially occurred for the IL32-beta transcript and corresponds with previous reports demonstrating that IL32-beta is not secreted from the cell like other canonical interleukins and preferentially localizes to the mitochondria in breast cancer cells. Additionally, expression of the beta-transcript could be suppressed when CSC-enriched cells were treated with the BET-bromodomain inhibitor JQ1. Because of this phenomenon, we sought to determine the effects of suppressing IL32 in the EpCAM-/CD49f- enriched cell line SUM159PT via siRNA-mediated knockdown and subsequent RNAseq differential expression analysis. From our results, we determined that transcripts involved in extracellular matrix (ECM) organization and collagen/integrin interaction were preferentially affected by IL32 silencing. Additionally, IL32 suppression decreased the invasiveness of SUM159PT based on an ECM-matrix cell invasion assay. Collectively, our results reflect the notion that differential IL32 expression by promoter hypomethylation in breast CSCs plays a role in ECM remodeling for purposes of breast cancer cell invasion and metastasis.

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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