寻常型银屑病患者皮损组织中DNA去甲基化酶TET3过表达介导转录因子KLF4显著高表达  

Overexpression of DNA demethylase ten-eleven translocation 3 mediates high expression of Krüppel-like factor 4 in the skin lesions of patients with psoriasis vulgaris

作　　者：张帅 罗雯 张莲 殷秀琴 罗宏 ZHANG Shuai;LUO Wen;ZHANG Lian;YIN Xiuqin;LUO Hong(Department of Dermatology,Changsha First Hospital,Changsha,Hunan Province,410000,China)

机构地区：[1]长沙市第一医院皮肤科,长沙410000

出　　处：《陆军军医大学学报》2024年第6期637-643,共7页Journal of Army Medical University

基　　金：湖南省卫生健康委科研计划(202204122971)。

摘　　要：目的 探讨银屑病患者皮损组织中Krüppel样因子4(Krüppel-like factor 4, KLF)异常高表达的DNA甲基化分子机制。方法 以20例寻常型银屑病患者病理检查所取皮损组织为实验组,13例眼外伤手术患者所取眼睑皮肤组织为对照组。采用亚硫酸氢盐测序(bisulfite sequencing, BSP)检测各组皮肤样本及转染十-十一转位3(ten-eleven translocation 3, TET3)过表达及干扰质粒的人永生化角质形成细胞HaCat中转录因子Krüppel样因子4(Krüppel-like factor 4, KLF4)启动子区DNA甲基化水平,RT-qPCR和Western blotting检测各组皮肤组织样本及转染HaCat细胞中KLF4、TET1(ten-eleven translocation1)、TET2、TET3的表达水平。结果 实验组KLF4启动子DNA甲基化水平显著低于对照组(P<0.01),实验组KLF4的mRNA表达水平显著高于对照组(P<0.01)。实验组KLF4启动子DNA甲基化水平与mRNA表达水平呈显著负相关(r=-0.649,P=0.002)。实验组TET3表达水平显著高于对照组(P<0.01), TET1及TET2表达水平差异无统计学意义。实验组TET3表达水平与KLF4启动子DNA甲基化水平呈显著负相关(r=-0.688,P=0.001)。HaCat细胞中过表达TET3后,KLF4启动子DNA甲基化水平明显下调(P<0.01),KLF4表达水平显著上调(P<0.01)。HaCat细胞中干扰TET3表达后,KLF4启动子DNA甲基化水平明显上调(P<0.01),KLF4表达水平显著下调(P<0.01)。结论 过度表达的TET3通过下调KLF4启动子DNA甲基化水平,介导银屑病患者皮损组织中KLF4过度表达。Objective To investigate the molecular mechanism of DNA methylation of the abnormally high expression of Krüppel-like factor 4(KLF4)in psoriasis vulgaris lesions.Methods The skin lesions of 20 patients with psoriasis vulgaris were taken as experimental group and those from 13 healthy individuals as control group.Bisulfite sequencing(BSP)was used to detect the DNA methylation of KLF4 promoter in the skin samples and in HaCat cells with ten-eleven translocation 3(TET3)overexpression and interference,respectively.Real time PCR and Western blotting were employed to detect the expression levels of KLF4,TET1,TET2 and TET3 in skin samples and transfected HaCat cells.Results The DNA methylation level of KLF4 promoter was significantly lower(P<0.01),and the KLF4 mRNA level was obviously higher in the experimental group than the control group(P<0.01).A negative correlation was observed between DNA methylation level of KLF4 promoter and KLF4 mRNA level in the experimental group(r=-0.649,P=0.002).The experimental group had notably higher expression level of TET3(P<0.01),but had no differences in the levels of TET1 and TET2 when compared with the control group.The expression level of TET3 was negatively correlated with the DNA methylation level of KLF4 promoter in the experimental group(r=-0.688,P=0.001).After overexpression of TET3 in HaCat cells,the DNA methylation level of KLF4 promoter was significantly decreased(P<0.01),and KLF4 expression level was obviously increased(P<0.01).After interfering with TET3 expression in HaCat cells,the DNA methylation level of KLF4 promoter was notably up-regulated(P<0.01),and the KLF4 expression level was remarkably decreased(P<0.01).Conclusion Overexpression of TET3 mediates the overexpression of KLF4 in psoriatic lesions by down-regulating the DNA methylation level of KLF4 promoter.

关 键 词：银屑病 角质形成细胞 DNA甲基化 TET家族蛋白3 kruppel样因子4 

分 类 号：R322.99[医药卫生—人体解剖和组织胚胎学] R394.3[医药卫生—基础医学] R758.63