Epigenetic changes in Basal Cell Carcinoma affect SHH and WNT signaling components

PLoS One. 2012;7(12):e51710. doi: 10.1371/journal.pone.0051710. Epub 2012 Dec 17.

Abstract

Background: The genetic background of Basal Cell Carcinoma (BCC) has been studied extensively, while its epigenetic makeup has received comparatively little attention. Epigenetic alterations such as promoter hypermethylation silence tumor suppressor genes (TSG) in several malignancies.

Objective: We sought to analyze the promoter methylation status of ten putative (tumor suppressor) genes that are associated with Sonic Hedgehog (SHH), WNT signaling and (hair follicle) tumors in a large series of 112 BCC and 124 healthy control samples by methylation-specific PCR.

Results: Gene promoters of SHH (P = 0.016), adenomatous polyposis coli (APC) (P = 0.003), secreted frizzled-related protein 5 (SFRP5) (P = 0.004) and Ras association domain family 1A (RASSF1A) (P = 0.023) showed significantly more methylation in BCC versus normal skin. mRNA levels of these four genes were reduced for APC and SFRP5 in BCC (n = 6) vs normal skin (n = 6). Down regulation of SHH, APC and RASSF1A could be confirmed on protein level as well (P<0.001 for all genes) by immunohistochemical staining. Increased canonical WNT activity was visualized by β-catenin staining, showing nuclear β-catenin in only 28/101 (27.7%) of BCC. Absence of nuclear β-catenin in some samples may be due to high levels of membranous E-cadherin (in 94.1% of the samples).

Conclusions: We provide evidence that promoter hypermethylation of key players within the SHH and WNT pathways is frequent in BCC, consistent with their known constitutive activation in BCC. Epigenetic gene silencing putatively contributes to BCC tumorigenesis, indicating new venues for treatment.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Carcinoma, Basal Cell / genetics*
  • Case-Control Studies
  • DNA Methylation*
  • Epigenesis, Genetic*
  • Female
  • Gene Expression Regulation, Neoplastic
  • Hedgehog Proteins / genetics*
  • Humans
  • Male
  • Promoter Regions, Genetic / genetics
  • Skin / metabolism*
  • Skin Neoplasms / genetics*
  • Tumor Cells, Cultured
  • Tumor Suppressor Proteins / genetics
  • Wnt Proteins / genetics*

Substances

  • Hedgehog Proteins
  • SHH protein, human
  • Tumor Suppressor Proteins
  • Wnt Proteins

Grants and funding

This work was supported by the Dutch Cancer Society, grants to TB (KWF UM2009-4609) and MVS (KWF UM2009-4352). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.