Luo Jingyi, Liu Tingting, Teng Weiping
Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Hospital of China Medical University, Shenyang, China.
J Mol Endocrinol. 2020 May;64(4):259-270. doi: 10.1530/JME-19-0239.
Hashimoto's thyroiditis (HT) is a common organ-specific autoimmune disease, which develops in 0.3-1.5/1000 subjects annually. The aims of this study were to determine the lncRNA profile in peripheral blood CD4+ T cells from HT patients and then to characterize the potential function of NONHSAT079547.2. A total of 37 HT patients and 50 sex- and age-matched healthy controls were enrolled for high-throughput sequencing. Another 43 HT patients and 50 sex- and age-matched controls were enrolled for validation via real-time PCR. Flow cytometry and CCK8 assays were used to measure cell apoptosis and growth levels. Western blotting was used for measuring the expression of growth- and apoptosis-associated proteins. IL-17 serum concentration and transcriptional level in CD4+ T cells of participants were detected by ELISA and real-time PCR, respectively. The mechanism of competitive endogenous RNA was determined using real-time PCR, ELISA, RNA immunoprecipitation, and dual-luciferase assays in Jurkat cells. A total of 7564 significantly differentially expressed lncRNAs were found, of which 3913 lncRNAs were upregulated and 3651 lncRNAs were downregulated in HT group when compared to control group. NONHSAT079547.2 was significantly upregulated in HT patients and was positively correlated with serum thyroid peroxidase antibody level. Further studies confirmed that NONHSAT079547.2 could promote cell growth and control IL-17 expression and secretion via the NONHSAT079547.2/miR-4716-5p/IL-17 axis.This is the first study to describe the lncRNA profile in CD4+ T cells of HT patients. The studies on the function of NONHSAT079547.2 might elucidate the underlying molecular mechanisms and represent potential biomarkers for HT.
桥本甲状腺炎(HT)是一种常见的器官特异性自身免疫性疾病,每年在每1000名受试者中发病率为0.3 - 1.5。本研究的目的是确定HT患者外周血CD4 + T细胞中的lncRNA谱,然后表征NONHSAT079547.2的潜在功能。共纳入37例HT患者和50例性别及年龄匹配的健康对照进行高通量测序。另外纳入43例HT患者和50例性别及年龄匹配的对照通过实时PCR进行验证。采用流式细胞术和CCK8法检测细胞凋亡和生长水平。蛋白质印迹法用于检测生长和凋亡相关蛋白的表达。分别通过ELISA和实时PCR检测参与者血清IL - 17浓度和CD4 + T细胞中的转录水平。在Jurkat细胞中使用实时PCR、ELISA、RNA免疫沉淀和双荧光素酶测定法确定竞争性内源RNA的机制。共发现7564个差异表达显著的lncRNA,与对照组相比,HT组中3913个lncRNA上调,3651个lncRNA下调。NONHSAT079547.2在HT患者中显著上调,且与血清甲状腺过氧化物酶抗体水平呈正相关。进一步研究证实,NONHSAT079547.2可通过NONHSAT079547.2/miR - 4716 - 5p/IL - 17轴促进细胞生长并调控IL - 17的表达和分泌。这是第一项描述HT患者CD4 + T细胞中lncRNA谱的研究。对NONHSAT079547.2功能的研究可能阐明潜在的分子机制,并代表HT的潜在生物标志物。