Black raspberries demethylate Sfrp4, a WNT pathway antagonist, in rat esophageal squamous cell papilloma

Mol Carcinog. 2016 Nov;55(11):1867-1875. doi: 10.1002/mc.22435. Epub 2015 Nov 19.


Aberrant methylation of DNA is a common event in the development of cancers, including squamous cell carcinoma (SCC) of the human esophagus. In the present study, we determined: (a) whether aberrant DNA methylation also occurs in the development of N-nitrosomethylbenzylamine (NMBA)-induced tumorigenesis in the rat esophagus, a model of human esophageal SCC; and (b) if so, whether dietary black raspberries (BRBs) are capable of preventing this aberrant DNA methylation. A diet containing 5% BRBs inhibited the development of NMBA-induced tumors in the rat esophagus. This inhibition was associated with reduced mRNA levels of the DNA methyltransferases, Dnmt1 and Dnmt3b, in both dysplastic lesions and in papillomas of the esophagus. In addition, promoter methylation of Sfrp4, a WNT pathway antagonist, was significantly reduced by the berry diet, and this was associated with decreased nuclear localization of β-CATENIN and reduced expression of c-MYC protein in NMBA-treated esophagi. Decreased promoter methylation of Sfrp4 correlated with decreased expression of Dmnt3b and, ultimately, with increased Sfrp4 mRNA expression. This suggests that epigenetic alterations in NMBA-induced rat esophageal tumorigenesis recapitulate epigenetic events in human esophageal SCC, and that BRBs could be useful in preventing the aberrant DNA methylation involved in the development of human esophageal SCC. © 2015 Wiley Periodicals, Inc.

Keywords: DNA methylation; Sfrp4; WNT signaling; black raspberries; rat esophageal tumor.

MeSH terms

  • Animals
  • Carcinoma, Squamous Cell / chemically induced
  • Carcinoma, Squamous Cell / diet therapy*
  • Carcinoma, Squamous Cell / genetics
  • DNA (Cytosine-5-)-Methyltransferase 1
  • DNA (Cytosine-5-)-Methyltransferases / genetics*
  • DNA Methylation / drug effects
  • DNA Methyltransferase 3B
  • Dimethylnitrosamine / adverse effects
  • Dimethylnitrosamine / analogs & derivatives*
  • Epigenesis, Genetic / drug effects
  • Esophageal Neoplasms / chemically induced
  • Esophageal Neoplasms / diet therapy*
  • Esophageal Neoplasms / genetics
  • Esophageal Squamous Cell Carcinoma
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Male
  • Plant Extracts / administration & dosage*
  • Plant Extracts / pharmacology
  • Proto-Oncogene Proteins / genetics*
  • Rats
  • Rubus / chemistry*
  • Wnt Signaling Pathway / drug effects


  • Plant Extracts
  • Proto-Oncogene Proteins
  • Sfrp4 protein, rat
  • nitrosobenzylmethylamine
  • DNA (Cytosine-5-)-Methyltransferase 1
  • DNA (Cytosine-5-)-Methyltransferases
  • DNMT1 protein, human
  • Dnmt1 protein, rat
  • Dimethylnitrosamine