title: Epigenetic silencing of DSC3 is a common event in human breast cancer.
abstract: BACKGROUND
  Desmocollin 3 (DSC3) is a member of the cadherin superfamily of calcium-dependent cell adhesion molecules and a principle component of desmosomes. Desmosomal proteins such as DSC3 are integral to the maintenance of tissue architecture and the loss of these components leads to a lack of adhesion and a gain of cellular mobility. DSC3 expression is down-regulated in breast cancer cell lines and primary breast tumors; however, the loss of DSC3 is not due to gene deletion or gross rearrangement of the gene. In this study, we examined the prevalence of epigenetic silencing of DSC3 gene expression in primary breast tumor specimens.
  
  
  METHODS
  We used bisulfite genomic sequencing to analyze the methylation state of the DSC3 promoter region from 32 primary breast tumor specimens. We also used a quantitative real-time RT-PCR approach, and analyzed all breast tumor specimens for DSC3 expression. Finally, in addition to bisulfite sequencing and RT-PCR, we used an in vivo nuclease accessibility assay to determine the chromatin architecture of the CpG island region from DSC3-negative breast cancer cells lines.
  
  
  RESULTS
  DSC3 expression was downregulated in 23 of 32 (72%) breast cancer specimens comprising: 22 invasive ductal carcinomas, 7 invasive lobular breast carcinomas, 2 invasive ductal carcinomas that metastasized to the lymph node, and a mucoid ductal carcinoma. Of the 23 specimens showing a loss of DSC3 expression, 13 (56%) were associated with cytosine hypermethylation of the promoter region. Furthermore, DSC3 expression is limited to cells of epithelial origin and its expression of mRNA and protein is lost in a high proportion of breast tumor cell lines (79%). Lastly, DNA hypermethylation of the DSC3 promoter is highly correlated with a closed chromatin structure.
  
  
  CONCLUSIONS
  These results indicate that the loss of DSC3 expression is a common event in primary breast tumor specimens, and that DSC3 gene silencing in breast tumors is frequently linked to aberrant cytosine methylation and concomitant changes in chromatin structure.
authors.affiliation: Department of Pharmacology, Arizona Cancer Center, University of Arizona, Tucson, AZ, USA. moshiro@azcc.arizona.edu
authors.forename: Marc M
authors.initials: MM
authors.lastname: Oshiro
authors.forename: Christina J
authors.initials: CJ
authors.lastname: Kim
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Wozniak
authors.forename: Damian J
authors.initials: DJ
authors.lastname: Junk
authors.forename: José L
authors.initials: JL
authors.lastname: Muñoz-Rodríguez
authors.forename: Jeanne A
authors.initials: JA
authors.lastname: Burr
authors.forename: Matthew
authors.initials: M
authors.lastname: Fitzgerald
authors.forename: Sangita C
authors.initials: SC
authors.lastname: Pawar
authors.forename: Anne E
authors.initials: AE
authors.lastname: Cress
authors.forename: Frederick E
authors.initials: FE
authors.lastname: Domann
authors.forename: Bernard W
authors.initials: BW
authors.lastname: Futscher
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D019175:DNA Methylation; D017931:DNA Primers; D051187:Desmocollins; D004847:Epithelial Cells / Q000201:enzymology; D005260:Female; D020868:Gene Silencing; D006801:Humans; D008297:Male; D008408:Mastectomy; D015412:Mastectomy, Segmental; D008562:Membrane Glycoproteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011467:Prostate / Q000201:enzymology; D012334:RNA, Neoplasm / Q000235:genetics
chemical_list: D017931:DNA Primers; C490356:DSC3 protein, human; D051187:Desmocollins; D008562:Membrane Glycoproteins; D012334:RNA, Neoplasm