title: Hypermethylation-Induced Inactivation of the IRF6 Gene as a Possible Early Event in Progression of Vulvar Squamous Cell Carcinoma Associated With Lichen Sclerosus.
abstract: The molecular mechanism leading to the development of vulvar squamous cell carcinoma (VSCC) from vulvar lichen sclerosus (VLS) is unknown.
  
  
  
  To assess the possible involvement of the IRF6 tumor-suppressor gene in the development of VSCC from VLS.
  
  
  
  In laboratories at the University of Ferrara, Ferrara, Italy, IRF6 gene expression and promoter methylation were investigated in paraffin-embedded VSCC and adjacent vulvar intraepithelial neoplasia (VIN) and VLS specimens, in cancer-free VLS (cfVLS) specimens, and in healthy skin specimens by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) analysis and by sequencing of PCR-amplified bisulfite-treated DNA. Methylation-induced dysregulation was assessed by expression of p63, the IRF6-transactivator gene.
  
  
  
  IRF6 expression, correlation with promoter methylation and p63 expression, and association with development of VSCC from VLS.
  
  
  
  Specimens from 60 participating women (1 specimen from each) were analyzed for the study (mean [SD] age, 66.3 [12.1] years): 20 paraffin-embedded specimens of VSCC (patient age, 75.3 [8.3] years) with adjacent VLS lesions, in 5 of which VIN preneoplastic tissue was also present (patient age, 64.3 [15.3] years); 20 cfVLS specimens (patient age, 62.1 [11.4] years) obtained from diagnostic biopsies; and 20 normal skin specimens from noncancer surgical patients (patient age, 61.4 [9.1] years). IRF6 messenger RNA was found to be reduced 4.5-, 2.9-, 6.6-, and 2.2-fold in VLS, VIN, VSCC, and cfVLS specimens, respectively, compared with controls, although p63 was still expressed in all specimens. IRF6 promoter was hypermethylated in 9 (45%) of 20 VLS specimens, 1 (20%) of 5 VIN specimens, 16 (80%) of 20 VSCC specimens, 2 (10%) of 20 cfVLS specimens, and 0 normal skin specimens.
  
  
  
  IRF6 dysregulation may be involved in the development of VSCC from VLS. Methylation of the IRF6 promoter may be a marker of cancer risk in patients with VLS.
authors.affiliation: Section of Pathology, Oncology, and Experimental Biology, Department of Morphology, Surgery, and Experimental Medicine, University of Ferrara, Ferrara, Italy.
authors.forename: John Charles
authors.initials: JC
authors.lastname: Rotondo
authors.affiliation: Section of Dermatology and Infectious Diseases, Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Borghi
authors.affiliation: Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Rita
authors.initials: R
authors.lastname: Selvatici
authors.affiliation: Section of Anatomic Pathology, Department of Morphology, Surgery, and Experimental Medicine, University of Ferrara and S. Anna University Hospital, Ferrara, Italy.
authors.forename: Eros
authors.initials: E
authors.lastname: Magri
authors.affiliation: Section of Anatomic Pathology, Department of Morphology, Surgery, and Experimental Medicine, University of Ferrara and S. Anna University Hospital, Ferrara, Italy.
authors.forename: Enzo
authors.initials: E
authors.lastname: Bianchini
authors.affiliation: Section of Anatomic Pathology, Department of Morphology, Surgery, and Experimental Medicine, University of Ferrara and S. Anna University Hospital, Ferrara, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Montinari
authors.affiliation: Section of Dermatology and Infectious Diseases, Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Corazza
authors.affiliation: Section of Dermatology and Infectious Diseases, Department of Medical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Annarosa
authors.initials: A
authors.lastname: Virgili
authors.affiliation: Section of Pathology, Oncology, and Experimental Biology, Department of Morphology, Surgery, and Experimental Medicine, University of Ferrara, Ferrara, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Tognon
authors.affiliation: Section of Pathology, Oncology, and Experimental Biology, Department of Morphology, Surgery, and Experimental Medicine, University of Ferrara, Ferrara, Italy.
authors.forename: Fernanda
authors.initials: F
authors.lastname: Martini
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002278:Carcinoma in Situ / Q000235:genetics*; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000473:pathology; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D019175:DNA Methylation; D005260:Female; D015870:Gene Expression; D006801:Humans; D050835:Interferon Regulatory Factors / Q000235:genetics*; D018459:Lichen Sclerosus et Atrophicus / Q000235:genetics* / Q000473:pathology; D008875:Middle Aged; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis*; D012867:Skin / Q000737:chemistry; D014157:Transcription Factors / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000235:genetics; D014846:Vulvar Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: C109803:IRF6 protein, human; D050835:Interferon Regulatory Factors; D012333:RNA, Messenger; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins