ABSTRACT

Psoriasis is an autoimmune skin disorder influenced by genetic, epigenetic and environmental factors. We previously found CYP2S1 intragenic DNA methylation cg19430423 site strongly hypomethylated in psoriatic skin tissues. In this study, we performed methylation loci fine-mapping to search the top signals in the entire CYP2S1 gene region, and further carried out gene expression assay, cell proliferation, apoptosis, differentiation and migration in CYP2S1 overexpressed (CYP2S1^over) and silenced (siRNA) human keratinocytes. Target bisulphite conversion sequencing revealed cg19430423 and nearby two loci were the top differentially methylated loci. These three loci located within active enhancer region marked by H3K4Me1 and H3K27AC peaks. Cg19430423 might not bind with ATF1 directly. CYP2S1^over repressed NHEK cell proliferation, but have no confirmed evidence on affecting migration, apoptosis and differentiation. Real-time PCR showed that CYP2S1 inhibited expression of IL1β, IL8, IL33, IL36, LL37, CXCL10 and CCL20 gene. In summary, CYP2S1 might inhibit keratinocyte proliferation, and modulate immune response through IL-8, IL-33, IL-36, CXCL-10, CCL20, thus contribute to the development of psoriasis.

摘要

银屑病是一种受遗传、表观遗传和环境因素影响的自身免疫性皮肤病。我们之前在银屑病皮肤组织中发现 CYP2S1基因内 DNA 甲基化 cg19430423 位点强烈低甲基化。在本研究中，我们进行甲基化位点精细定位以搜索整个 CYP2S1 基因区域的顶部信号，并进一步进行基因表达测定，  CYP2S1 过表达（CYP2S1 ^over) 和沉默 (siRNA) 人角质形成细胞。目标亚硫酸氢盐转化测序显示 cg19430423 和附近的两个基因座是顶部差异甲基化基因座。这三个基因座位于由 H3K4Me1 和 H3K27AC 峰标记的活性增强子区域内。Cg19430423 可能不直接与 ATF1 结合。 CYP2S1^过度 抑制 NHEK 细胞增殖，但在影响迁移、细胞凋亡和分化方面没有确凿的证据。实时荧光定量 PCR 显示 CYP2S1 抑制 IL1β、IL8、IL33、IL36、LL37、CXCL10 和 CCL20 基因的表达。总之，  CYP2S1可能抑制角质形成细胞增殖，并通过IL-8、IL-33、IL-36、CXCL-10、CCL20 调节免疫反应 ，从而有助于牛皮癣的发展。