Functional characterization of recurrent FOXA2 mutations seen in endometrial cancers

Int J Cancer. 2018 Dec 1;143(11):2955-2961. doi: 10.1002/ijc.31784. Epub 2018 Sep 29.

Abstract

FOXA2, a member of the forkhead family of DNA-binding proteins, is frequently mutated in uterine cancers. Most of the mutations observed in uterine cancers are frameshifts and stops. FOXA2 is considered to be a driver gene in uterine cancers, functioning as a haploinsufficient tumor suppressor. The functional consequences of FOXA2 mutations, however, have not yet been determined. We evaluated the effects that frameshift mutations and a recurrent missense mutation have on FOXA2 transcriptional activity. Recurrent N-terminal frameshifts resulted in truncated proteins that failed to translocate to the nucleus and have no transcriptional activity using an E-cadherin/luciferase reporter assay. Protein abundance was reduced for the recurrent p.S169 W mutation, as was transcriptional activity. A C-terminal frameshift mutation had increased FOXA2 levels evidenced by both Western blot and immunofluorescence. Given that FOXA2 is a recognized activator of E-cadherin (CDH1) expression and E-cadherin's potential role in epithelial-to-mesenchymal transition in a wide range of cancer types, we tested the hypothesis that FOXA2 mutations in primary uterine cancer specimens would be associated with reduced CDH1 transcript levels. qRT-PCR revealed significantly lower levels of CDH1 expression in primary tumors with FOXA2 mutations. Our findings in vitro and in vivo suggest that reduced transcriptional activity associated with FOXA2 mutations in uterine cancers is likely to contribute to protumorigenic changes in gene expression.

Keywords: endometrial cancer; forkhead box protein FOXA2; tumor suppressor.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cadherins / genetics
  • Cell Line
  • Cell Line, Tumor
  • DNA-Binding Proteins / genetics
  • Endometrial Neoplasms / genetics*
  • Female
  • HEK293 Cells
  • Hepatocyte Nuclear Factor 3-beta / genetics*
  • Humans
  • Mutation / genetics*
  • Protein Transport / genetics
  • Transcription, Genetic / genetics

Substances

  • Cadherins
  • DNA-Binding Proteins
  • FOXA2 protein, human
  • Hepatocyte Nuclear Factor 3-beta