Karami J, Mahmoudi M, Amirzargar A, Gharshasbi M, Jamshidi A, Aslani S, Nicknam M H
Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Genes Immun. 2017 Sep;18(3):170-175. doi: 10.1038/gene.2017.17. Epub 2017 Aug 10.
Methylation of DNA is one of the important regulatory mechanisms of gene transcription. B-cell chronic lymphocytic leukemia/lymphoma 11B (BCL11B) plays a key role in the development, proliferation, differentiation, and survival of T cells. The aim of this study was to evaluate promoter methylation of BCL11B gene and its mRNA level in peripheral blood mononuclear cells (PBMCs) of ankylosing spondylitis (AS) patients in relation to healthy controls and evaluate their correlation with diseases clinical indices. Fifty AS patients and 50 healthy controls entered in this study. PBMCs were isolated from whole blood and RNA and DNA contents of leukocytes were extracted. The expression level of BCL11B gene was measured through real-time PCR assay using SYBR Green Master Mix, and PCR products of bisulfite-treated DNA were sequenced to determine the methylation level of promoter. Decreased transcript and increased promoter methylation levels of BCL11B gene were identified in AS patients compared with healthy controls. Hypermethylation of CpG3 and CpG5 was associated with increased AS risk. Promoter hypermethylation and mRNA overexpression correlated with each other but not with clinical manifestations. We identified aberrancies in expression and methylation of BCL11B gene in AS patients compared with healthy control group; however, they might not impress the clinical face of AS.
DNA甲基化是基因转录的重要调控机制之一。B细胞慢性淋巴细胞白血病/淋巴瘤11B（BCL11B）在T细胞的发育、增殖、分化和存活中起关键作用。本研究旨在评估强直性脊柱炎（AS）患者外周血单个核细胞（PBMC）中BCL11B基因的启动子甲基化及其mRNA水平，并与健康对照进行比较，同时评估它们与疾病临床指标的相关性。50例AS患者和50例健康对照纳入本研究。从全血中分离出PBMC，并提取白细胞的RNA和DNA。使用SYBR Green Master Mix通过实时PCR测定法测量BCL11B基因的表达水平，对亚硫酸氢盐处理后的DNA的PCR产物进行测序以确定启动子的甲基化水平。与健康对照相比，AS患者中BCL11B基因的转录本减少，启动子甲基化水平增加。CpG3和CpG5的高甲基化与AS风险增加相关。启动子高甲基化和mRNA过表达相互关联，但与临床表现无关。与健康对照组相比，我们在AS患者中发现了BCL11B基因表达和甲基化的异常；然而，它们可能不会影响AS的临床症状。
