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. 2008 May;44(7):1030-8.
doi: 10.1016/j.ejca.2008.02.046. Epub 2008 Apr 18.

Lineage-specific silencing of human IL-10 gene expression by promoter methylation in cervical cancer cells

Anita Szalmás  1 , Ferenc Bánáti, Anita Koroknai, Brigitta László, Eniko Fehér, Dániel Salamon, Lajos Gergely, János Minárovits, József Kónya
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Lineage-specific silencing of human IL-10 gene expression by promoter methylation in cervical cancer cells

Anita Szalmás et al. Eur J Cancer. 2008 May.

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.

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