Multifactorial regulation of E-cadherin expression: an integrative study

Mol Cancer Ther. 2010 Jan;9(1):1-16. doi: 10.1158/1535-7163.MCT-09-0321. Epub 2010 Jan 6.


E-cadherin (E-cad) is an adhesion molecule associated with tumor invasion and metastasis. Its down-regulation is associated with poor prognosis for many epithelial tumor types. We have profiled E-cad in the NCI-60 cancer cell lines at the DNA, RNA, and protein levels using six different microarray platforms plus bisulfite sequencing. Here we consider the effects on E-cad expression of eight potential regulatory factors: E-cad promoter DNA methylation, the transcript levels of six transcriptional repressors (SNAI1, SNAI2, TCF3, TCF8, TWIST1, and ZFHX1B), and E-cad DNA copy number. Combined bioinformatic and pharmacological analyses indicate the following ranking of influence on E-cad expression: (1) E-cad promoter methylation appears predominant, is strongly correlated with E-cad expression, and shows a 20% to 30% threshold above which E-cad expression is silenced; (2) TCF8 expression levels correlate with (-0.62) and predict (P < 0.00001) E-cad expression; (3) SNAI2 and ZFHX1B expression levels correlate positively with each other (+0.83) and also correlate with (-0.32 and -0.30, respectively) and predict (P = 0.03 and 0.01, respectively) E-cad expression; (4) TWIST1 correlates with (-0.34) but does not predict E-cad expression; and (5) SNAI1 expression, TCF3 expression, and E-cad DNA copy number do not correlate with or predict E-cad expression. Predictions of E-cad regulation based on the above factors were tested and verified by demethylation studies using 5-aza-2'-deoxycytidine treatment; siRNA knock-down of TCF8, SNAI2, or ZFHX1B expression; and combined treatment with 5-aza-2'-deoxycytidine and TCF8 siRNA. Finally, levels of cellular E-cad expression are associated with levels of cell-cell adhesion and response to drug treatment.

Publication types

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

MeSH terms

  • Azacitidine / pharmacology
  • Branched DNA Signal Amplification Assay
  • Cadherins / genetics*
  • Cadherins / metabolism
  • Cell Adhesion / drug effects
  • Cell Line, Tumor
  • Drug Screening Assays, Antitumor
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Gene Dosage / drug effects
  • Gene Expression Regulation, Neoplastic* / drug effects
  • Homeodomain Proteins / metabolism
  • Humans
  • Methylation / drug effects
  • Oligonucleotide Array Sequence Analysis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Transcription Factors / metabolism
  • Up-Regulation / drug effects
  • Zinc Finger E-box-Binding Homeobox 1


  • Cadherins
  • Homeodomain Proteins
  • RNA, Messenger
  • RNA, Small Interfering
  • Trans-Activators
  • Transcription Factors
  • ZEB1 protein, human
  • Zinc Finger E-box-Binding Homeobox 1
  • Azacitidine