title: Lineage-specific silencing of human IL-10 gene expression by promoter methylation in cervical cancer cells.
abstract: Epigenetic analysis was performed to demonstrate that the normal and neoplastic epithelial cells do not serve as the source of the locally elevated IL-10 production during cervical carcinogenesis. Bisulfite sequencing was used to correlate promoter CpG methylation with the transcription of the gene. Lack of IL-10 transcription in HeLa, SiHa, Caski, HT-3, C33-A, HaCaT cell lines and in primary human keratinocytes correlated consistently with the methylated state of the proximal CpG residues, particularly with the two most proximal CpGs at positions -185 and -110. These two sites were also highly methylated in normal and malignant cervical cells directly isolated from patient material. On the other hand, IL-10 producing peripheral blood mononuclear cells had unmethylated CpG residues in the proximal promoter associated with acetylated H3 and H4 histones as determined by chromatin immunoprecipitation. In HeLa carrying epigenetically silenced endogeneous IL-10 promoter, the transfected non-CpG methylated 1 kb and 0.6 kb proximal promoter fragments could drive reporter gene expression, which was reversed by cassette methylation of these promoter fragments. In conclusion, the CpG methylation pattern of the proximal promoter is implicated as a major determinant of transcriptional silencing of human IL-10 expression in cells of cervical epithelial origin.
authors: Department of Medical Microbiology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary.
authors: Anita
authors: A
authors: Szalmás
authors: Ferenc
authors: F
authors: Bánáti
authors: Anita
authors: A
authors: Koroknai
authors: Brigitta
authors: B
authors: László
authors: Eniko
authors: E
authors: Fehér
authors: Dániel
authors: D
authors: Salamon
authors: Lajos
authors: L
authors: Gergely
authors: János
authors: J
authors: Minárovits
authors: József
authors: J
authors: Kónya
mesh_terms: D045744:Cell Line, Tumor; D019175:DNA Methylation; D005260:Female; D015870:Gene Expression / Q000235:genetics; D020868:Gene Silencing; D051548:Histone Acetyltransferases / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics*; D015603:Keratinocytes / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D002583:Uterine Cervical Neoplasms / Q000235:genetics*
chemical_list: D016753:Interleukin-10; D051548:Histone Acetyltransferases