Lu Ming-Chi, Yu Hui-Chun, Yu Chia-Li, Huang Hsien-Bin, Koo Malcolm, Tung Chien-Hsueh, Lai Ning-Sheng
Division of Allergy, Immunology and Rheumatology, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, No. 2, Minsheng Road, Dalin, Chiayi, 62247, Taiwan.
School of Medicine, Tzu Chi University, Hualien, Taiwan.
Immunol Res. 2016 Apr;64(2):576-83. doi: 10.1007/s12026-015-8756-8.
The aim of this study was to evaluate whether the presence of aberrantly expressed lncRNAs could promote T cell inflammatory responses in patients with RA. The expression levels of 10 potential aberrantly expressed lncRNAs were evaluated in T cells from 39 patients with RA and 17 controls using real-time reverse transcription polymerase chain reaction. The aberrantly expressed lncRNAs were measured in Jurkat cells co-cultured with or without ionomycin and phorbol 12-myristate 13-acetate. Transfection studies using small interfering RNA (siRNA) were conducted for biological functions, and microarray analysis was performed to search for target genes of specific lncRNAs. We confirmed that the expression levels of LOC100652951 and LOC100506036 were higher in RA T cells compared with controls. RA patients treated with biologic agents had lower expression levels of LOC100652951, and female RA patients had lower LOC100506036 expression levels after multivariate analysis. After activation, the expression levels of LOC100506036, but not LOC100652951, increased in Jurkat cells. Transfection of siRNA targeting LOC100506036 inhibited interferon gamma production and the expression of nuclear factor of activated T cells in activated Jurkat cells. After the microarray analysis with validation, inhibition of LOC100506036 expression by siRNA leaded to the decreased expression of sphingomyelin phosphodiesterase 1 (SMPD1). In conclusion, the expression levels of LOC100652951 and LOC100506036 were increased in RA T cells. Treatment with biologic agents could lower the expression of LOC100652951 in RA T cells. LOC100506036 could regulate the expression of SMPD1 and NFAT1 and could contribute to the inflammatory responses in RA.
本研究的目的是评估异常表达的长链非编码RNA（lncRNAs）是否能促进类风湿关节炎（RA）患者的T细胞炎症反应。使用实时逆转录聚合酶链反应评估了39例RA患者和17例对照的T细胞中10种潜在异常表达的lncRNAs的表达水平。在与离子霉素和佛波酯12-肉豆蔻酸酯13-乙酸酯共培养或未共培养的Jurkat细胞中测量异常表达的lncRNAs。使用小干扰RNA（siRNA）进行转染研究以探讨生物学功能，并进行微阵列分析以寻找特定lncRNAs的靶基因。我们证实，与对照组相比，RA患者T细胞中LOC100652951和LOC100506036的表达水平更高。经多变量分析，接受生物制剂治疗的RA患者中LOC100652951的表达水平较低，女性RA患者中LOC100506036的表达水平较低。激活后，Jurkat细胞中LOC100506036的表达水平升高，而LOC100652951的表达水平未升高。靶向LOC100506036的siRNA转染抑制了活化的Jurkat细胞中γ干扰素的产生和活化T细胞核因子的表达。经过验证的微阵列分析后，siRNA抑制LOC100506036的表达导致鞘磷脂磷酸二酯酶1（SMPD1）的表达降低。总之，RA患者T细胞中LOC100652951和LOC100506036的表达水平升高。生物制剂治疗可降低RA患者T细胞中LOC100652951的表达。LOC100506036可调节SMPD1和NFAT1的表达，并可能参与RA的炎症反应。
