Changes in the DNA Methylation and Hydroxymethylation Status of the Intercellular Adhesion Molecule 1 Gene Promoter in Thyrocytes from Autoimmune Thyroiditis Patients
Changes in the DNA Methylation and Hydroxymethylation Status of the Intercellular Adhesion Molecule 1 Gene Promoter in Thyrocytes from Autoimmune Thyroiditis Patients
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自身免疫性甲状腺炎患者甲状腺细胞细胞间粘附分子1基因启动子DNA甲基化和羟甲基化状态的变化
DOI：
10.1089/thy.2016.0576
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发表时间：
2017
期刊：
影响因子：
6.6
通讯作者：
Teng Weiping
中科院分区：
文献类型：
--
作者：
Liu Tingting;Sun Jie;Wang Zhaojun;Yang Wenqing;Zhang Hao;Fan Chenling;Shan Zhongyan;Teng Weiping
背景：细胞间粘附分子1 （ICAM1）基因在正常甲状腺组织中不表达，但在自身免疫性甲状腺炎（AIT）患者的甲状腺组织中表达。方法：本研究旨在探讨在AIT患者甲状腺细胞中，icam1启动子的DNA甲基化和羟甲基化状态是否异常改变，以及这种改变是否与icam1的功能失调表达有关。共有35名AIT患者和35名性别和年龄匹配的对照组进行了研究。密度梯度离心分离甲状腺细胞后，实时荧光定量PCR检测ICAM1mRNA表达。在T4 β-葡萄糖基转移酶处理和mspi /HpaII在转录起始位点（TSS）上游- 937 bp、- 701 bp、- 226 bp和- 65 bp切割后，使用定量PCR评估DNA甲基化和羟甲基化状态。通过焦磷酸测序验证DNA甲基化水平。结果：AIT组在TSS上游- 937 bp、- 226 bp处DNA羟甲基化升高，在TSS上游- 937 bp、- 701 bp、- 226 bp处甲基化降低。焦磷酸测序还显示TSS上游- 708 bp、- 692 bp、- 690 bp和- 688 bp处的DNA低甲基化。TSS上游- 708 bp、- 692 bp和- 226 bp的DNA甲基化状态与icam1mrna的表达呈负相关。结论：总之，我们在AIT患者的甲状腺细胞中发现了异常的icam1基因启动子DNA甲基化和羟甲基化。这种异常的表观遗传修饰与icam1基因的表达增加有关。
Background:The intercellular adhesion molecule 1 (ICAM1) gene is not expressed in normal thyroid tissue but was shown to be expressed in the thyroid tissue of autoimmune thyroiditis (AIT) patients.Methods:This study aimed to explore whether the DNA methylation and hydroxymethylation status of theICAM1promoter are aberrantly altered in the thyroid cells of AIT patients and whether this change is associated with dysfunctional expression ofICAM1. A total of 35 AIT patients and 35 sex- and age-matched controls were studied. After the isolation of thyrocytes via density-gradient centrifugation,ICAM1mRNA expression was measured using real-time PCR. The DNA methylation and hydroxymethylation status were assessed using quantitative PCR following T4 β-glucosyltransferase treatment andMspI/HpaII cleavage at −937 bp, −701 bp, −226 bp, and −65 bp upstream of the transcription start site (TSS). The DNA methylation level was verified via pyrosequencing.Results:The AIT group showed increased DNA hydroxymethylation at −937 bp and −226 bp and decreased methylation at −937 bp, −701 bp, and −226 bp upstream of the TSS. Pyrosequencing also revealed DNA hypomethylation at −708 bp, −692 bp, −690 bp, and −688 bp upstream of the TSS. The DNA methylation status at −708 bp, −692 bp, and −226 bp upstream of the TSS was negatively associated withICAM1mRNA expression.Conclusion:In summary, we identified aberrant DNA methylation and hydroxymethylation of theICAM1gene promoter in the thyrocytes of AIT patients. This aberrant epigenetic modification is associated with increased expression of theICAM1gene.