ABSTRACT
OBJECTIVE: The purpose of this study was to discuss our investigation of the hypermethylation of promoter regions of tumor suppressor genes, such as death-associated protein kinase (DAPK) and p16, in vulvar lichen sclerosus (LS), in comparison with a control group. MATERIALS AND METHODS: Promoter hypermethylation of DAPK and p16 was investigated using 24 vulvar biopsies of patients with LS who had received no previous treatment. The control group was composed of 15 patients with no vulvar disease. The DNA of subjects was treated with sodium bisulphate, and the genes under study were subjected to methylation-specific polymerase chain reaction. The resulting polymerase chain reaction products were amplified and analyzed using a 10% polyacrylamide gel. RESULTS: The mean age of the patients with LS was 57 years (the majority were postmenopausal). In the control group, the mean age of the patients was 50 years (p = .151). Methylation of the promoter region of DAPK was found in 4 (17%) of the 23 patients analyzed, and p16 promoter region methylation was found in 8 patients (35%). Two cases of methylation of the DAPK gene were also found to be methylated for the p16 gene. In the control group, no methylation was found in the patients analyzed for the DAPK gene and methylation was found in 3 (21%) of the 14 patients analyzed for the p16 gene (p = .190 and p = .316, respectively). CONCLUSIONS: Methylation of the DAPK and p16 genes, although not sufficient to dictate prognosis of the disease, should not be underestimated because it may form part of a process of genetic and epigenetic alterations that in the future could become relevant to malignant transformation.
Subject(s)
Apoptosis Regulatory Proteins/genetics , Calcium-Calmodulin-Dependent Protein Kinases/genetics , DNA Methylation , Neoplasm Proteins/genetics , Promoter Regions, Genetic , Vulvar Lichen Sclerosus/genetics , Cyclin-Dependent Kinase Inhibitor p16 , Death-Associated Protein Kinases , Electrophoresis, Polyacrylamide Gel , Female , Humans , Middle Aged , Polymerase Chain Reaction/methods , Prognosis , Vulva/pathology , Vulvar Lichen Sclerosus/diagnosis , Vulvar Lichen Sclerosus/pathologyABSTRACT
OBJECTIVE: This article aimed to investigate the hypermethylation of promoter regions of tumor suppressor genes, such as death-associated protein kinase (DAPK) and p16, in vulvar lichen sclerosus (LS). MATERIALS AND METHODS: The promoter hypermethylation of DAPK and p16 was investigated from 15 vulvar biopsies of patients with LS who had had no previous treatment. DNA was treated with sodium bisulfate and underwent methylation-specific polymerase chain reaction of these genes. The amplified polymerase chain reaction products were analyzed by 10% polyacrylamide gel. RESULTS: The mean age of the patients was 57 years (most were postmenopausal). Methylation of the promoter region of DAPK was found in 2 (13%) of 15 patients analyzed, and p16 promoter region methylation was found in 7 patients (47%). The samples that showed DAPK methylation also showed p16 methylation. CONCLUSIONS: Methylation of DAPK and p16 represent alterations that might occur in cell cycle control in LS. The hypothesis is that patients who had methylated genes in this study, mainly the 2 cases in which there has been methylation in both studied genes, may be more susceptible to the development of differentiated vulvar intraepithelial neoplasia or vulvar cancer. Methylation may play a role in progress of vulvar carcinogenesis.
Subject(s)
Apoptosis Regulatory Proteins/genetics , Calcium-Calmodulin-Dependent Protein Kinases/genetics , DNA Methylation , DNA/metabolism , Neoplasm Proteins/genetics , Promoter Regions, Genetic , Vulvar Lichen Sclerosus/pathology , Cyclin-Dependent Kinase Inhibitor p16 , Death-Associated Protein Kinases , Electrophoresis, Polyacrylamide Gel/methods , Female , Humans , Middle Aged , Pathology, Molecular/methods , Polymerase Chain Reaction/methodsABSTRACT
OBJECTIVE: We investigated the presence of the Epstein-Barr virus (EBV) and human papillomavirus (HPV) in patients with vulvar lichen sclerosus (LS). MATERIALS AND METHODS: We investigated the presence of HPV and EBV from 34 vulvar biopsies of patients with LS who had had no previous treatment and from 17 normal vulvar brushings used as controls. We used polymerase chain reaction to amplify DNA sequences of these viruses. Human papillomavirus and EBV DNA detection was carried out using MY09/MY11 and TC67/TC69 consensus primers, respectively. The amplified polymerase chain reaction products were analyzed by 10% polyacrylamide gel. RESULTS: The mean age of the patients was 57 years old, with the majority postmenopausal. Human papillomavirus DNA was not found in the LS samples studied, but it was found in 23.2% (4/17) of the controls. However, EBV DNA was found in 26.5% (9/34) of the LS samples analyzed, and it was not found in the controls. CONCLUSIONS: Our results showed no relationship between HPV and LS. This result is in accordance with the literature. We have found 26.5% of EBV in our samples. This is a preliminary study, and the follow-up of these patients will elucidate whether EBV could play a role in cases of LS.
