Inhibition of CCN6 (Wnt-1-induced signaling protein 3) down-regulates E-cadherin in the breast epithelium through induction of snail and ZEB1

Am J Pathol. 2008 Apr;172(4):893-904. doi: 10.2353/ajpath.2008.070899. Epub 2008 Mar 5.

Abstract

The cysteine-rich protein CCN6 [or Wnt-1-induced signaling protein 3 (WISP3)] exerts tumor-suppressive effects in aggressive inflammatory breast cancer. Loss of CCN6 is associated with poorly differentiated phenotypes and increased invasion. Here, we show that reduction of CCN6 expression occurs in 60% of invasive breast carcinomas and is associated with axillary lymph node metastases. Furthermore, low CCN6 expression in invasive carcinoma tissue samples correlates with reduced expression of E-cadherin. In vitro, RNA interference knockdown of CCN6 in two benign human mammary epithelial cell lines (HME and MCF10A) decreased expression of E-cadherin protein and mRNA and reduced activity of the E-cadherin promoter; this reduction was dependent on intact E-box elements. CCN6 knockdown in HME cells resulted in up-regulation of the E-cadherin transcriptional repressors Snail and ZEB1 and enhanced their recruitment and binding to the E-cadherin promoter as analyzed by chromatin immunoprecipitation assays. Small interfering RNA-mediated knockdown of ZEB1 or Snail blocked the down-regulation of E-cadherin caused by CCN6 inhibition. These data show, for the first time, that CCN6 expression is reduced or lost in a substantial number of invasive breast carcinomas and that CCN6 modulates transcriptional repressors of E-cadherin. Together, our results lead to a new hypothesis that Snail and ZEB1 are downstream of CCN6 and play a critical role in CCN6-mediated regulation of E-cadherin in breast cancer.

Publication types

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

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Breast Neoplasms / genetics
  • Breast Neoplasms / pathology
  • CCN Intercellular Signaling Proteins
  • Cadherins / genetics*
  • Cadherins / metabolism
  • Cell Line, Tumor
  • Down-Regulation / genetics*
  • E-Box Elements / genetics
  • Epithelium / metabolism*
  • Epithelium / pathology
  • Female
  • Gene Expression Regulation, Neoplastic
  • Homeodomain Proteins / metabolism*
  • Humans
  • Insulin-Like Growth Factor Binding Proteins / metabolism*
  • Lymphatic Metastasis
  • Mammary Glands, Human / metabolism*
  • Mammary Glands, Human / pathology
  • Middle Aged
  • Neoplasm Invasiveness
  • Promoter Regions, Genetic / genetics
  • Protein Binding
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Repressor Proteins / metabolism
  • Snail Family Transcription Factors
  • Transcription Factors / metabolism*
  • Transcription, Genetic
  • Up-Regulation / genetics
  • Zinc Finger E-box-Binding Homeobox 1

Substances

  • CCN Intercellular Signaling Proteins
  • CCN6 protein, human
  • Cadherins
  • Homeodomain Proteins
  • Insulin-Like Growth Factor Binding Proteins
  • RNA, Messenger
  • Repressor Proteins
  • Snail Family Transcription Factors
  • Transcription Factors
  • ZEB1 protein, human
  • Zinc Finger E-box-Binding Homeobox 1