Molecular mechanism of transcriptional repression of gelsolin in human breast cancer cells

Exp Cell Res. 2002 Jun 10;276(2):328-36. doi: 10.1006/excr.2002.5534.

Abstract

Loss of gelsolin, a tumor suppressor, is one of the most frequently occurring molecular defects in breast cancers of diverse etiologies and across at least three animal species: human, mouse, and rat. Our previous analysis of breast cancer cells demonstrated that the deficiency is not due to mutation of the gelsolin gene, but instead to epigenetic factors, including decreased transcription of the gene. The study described herein provides the first functional characterization of the human gelsolin promoter and reveals a mechanistic basis for the reduced gelsolin transcription. In reporter gene assays, the gelsolin promoter was less active in low-gelsolin-expressing breast cancer cells. A cis-element mediating this reduced promoter activity was defined as a 27-bp sequence located approximately 135 bp upstream of the transcription start site. Gel shift and supershift assays and Southwestern blotting analysis indicated that activating transcription factor-1 (ATF-1) and a protein of approximately 100 kDa may have cancer cell-specific DNA-binding activity to the 27-bp gelsolin cis-element. Although the ATF-1 protein was highly expressed in both benign and tumorigenic breast cells, its DNA-binding activity was selectively abundant in the cancer cells and correlated inversely with the gelsolin mRNA level. Thus, our results suggest a role for ATF-1 in gelsolin promoter silencing in contrast to its transactivating effect on various other promoters.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 5' Flanking Region / genetics
  • Activating Transcription Factor 1
  • Base Sequence / genetics
  • Binding Sites / genetics
  • Breast / cytology
  • Breast / metabolism
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / physiopathology
  • Carcinoma / genetics*
  • Carcinoma / metabolism
  • Carcinoma / physiopathology
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Down-Regulation / genetics*
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism
  • Female
  • Gelsolin / deficiency
  • Gelsolin / genetics*
  • Genes, Regulator / genetics*
  • Humans
  • Molecular Sequence Data
  • Promoter Regions, Genetic / genetics
  • Protein Binding / physiology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Repressor Proteins / genetics*
  • Repressor Proteins / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / isolation & purification
  • Transcription Factors / metabolism
  • Tumor Cells, Cultured
  • Tumor Suppressor Proteins / deficiency
  • Tumor Suppressor Proteins / genetics*

Substances

  • Activating Transcription Factor 1
  • DNA-Binding Proteins
  • Gelsolin
  • RNA, Messenger
  • Repressor Proteins
  • Transcription Factors
  • Tumor Suppressor Proteins