Department of Epidemiology and Biostatistics, School of Public Health, Anhui Medical University, Hefei, China.
The Key Laboratory of Major Autoimmune Diseases, Anhui Medical University, Hefei, China.
Int J Rheum Dis. 2023 Dec;26(12):2526-2533. doi: 10.1111/1756-185X.14955. Epub 2023 Oct 30.
Ankylosing spondylitis is a refractory immune disease that seriously affects the life and work of patients. Epigenetic modifications, especially DNA methylation, have become a research hotspot in complex diseases. We aim to explore the changes in runt-related transcription factor 2 (RUNX2) gene promoter methylation and transcription level in AS.
We detected the RUNX2 gene promoter methylation in 83 AS patients and 83 healthy controls (HCs), then inspected the mRNA difference of RUNX2 between 30 AS patients and 30 HCs by the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR).
The RUNX2 gene promoter was hypomethylated in AS patients compared to HCs (p < .001). The research involved 4 CpG regions and 74 CpG sites of RUNX2, of which CpG-2, CpG-4 regions, and 18 CpG sites have been differentially methylated. The CpG-4 island methylation was negatively correlated with C-reactive protein (p < .05) in AS patients. In the qRT-PCR validation phase, the mRNA level of RUNX2 in AS patients was significantly higher than HCs (p < .05), and in AS patients who were treated with biologics, the methylation level of CpG-2 island showed a negative correlation to mRNA (p < .05). ROC results indicated that RUNX2 methylation and its transcription level have good potential to distinguish AS patients from HCs.
The RUNX2 gene promoter was hypomethylated in AS patients. Meanwhile, the qRT-PCR verified the up-regulated expression on the transcription level, suggesting the abnormal methylation of RUNX2 contributes to the pathogenesis of AS.
强直性脊柱炎是一种难治性免疫疾病，严重影响患者的生活和工作。表观遗传修饰，尤其是 DNA 甲基化，已成为复杂疾病的研究热点。我们旨在探讨强直性脊柱炎中 runt 相关转录因子 2（RUNX2）基因启动子甲基化和转录水平的变化。
我们检测了 83 例强直性脊柱炎患者和 83 例健康对照者（HCs）的 RUNX2 基因启动子甲基化情况，然后通过实时定量逆转录聚合酶链反应（qRT-PCR）检测了 30 例强直性脊柱炎患者和 30 例 HCs 之间 RUNX2 的 mRNA 差异。
与 HCs 相比，AS 患者的 RUNX2 基因启动子呈低甲基化状态（p<0.001）。该研究涉及 RUNX2 的 4 个 CpG 区和 74 个 CpG 位点，其中 CpG-2、CpG-4 区和 18 个 CpG 位点存在差异甲基化。AS 患者的 CpG-4 岛甲基化与 C-反应蛋白呈负相关（p<0.05）。在 qRT-PCR 验证阶段，AS 患者的 RUNX2 mRNA 水平明显高于 HCs（p<0.05），且在接受生物制剂治疗的 AS 患者中，CpG-2 岛的甲基化水平与 mRNA 呈负相关（p<0.05）。ROC 结果表明，RUNX2 甲基化及其转录水平具有很好的潜力将 AS 患者与 HCs 区分开来。
AS 患者的 RUNX2 基因启动子呈低甲基化状态。同时，qRT-PCR 验证了转录水平的上调表达，提示 RUNX2 的异常甲基化可能与 AS 的发病机制有关。
