Qiu Yu-Ying, Zhang Ying-Wei, Qian Xiu-Fen, Bian Tao
Department of Respiratory Medicine, The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNo. 321 Zhongshan Road, Nanjing 210008, Jiangsu, China.
Department of Respiratory Medicine, Wuxi People's Hospital Affiliated to Nanjing Medical UniversityNo. 299 Qingyang Road, Wuxi 214023, Jiangsu, China.
Am J Transl Res. 2017 Jul 15;9(7):3184-3199. eCollection 2017.
Asthma is tightly related to the imbalance of Th1/Th2 cells, and Runx3 plays a pivotal role in the differentiation of T helper cells. The present study aimed to investigate dysregulated microRNAs that may target Runx3 in CD4 T cells from asthmatic patients and reveal Runx3 function in Th1/Th2 balance regulation. We detected the levels of Th1- and Th2-related cytokines by ELISA and analyzed the differentiation marker gene of T helper cells by qRT-PCR. Results indicated that an imbalance of Th1/Th2 cells was present in our asthmatic subject. Runx3 expression was reduced in the CD4 T cells from asthmatic patients. Overexpression of Runx3 could restore the Th1/Th2 balance. After performing microRNA microarray assay, we found a series of microRNAs that were considerably altered in the CD4 T cells from asthmatic patients. Among these upregulated microRNAs, eight microRNAs that may target Runx3 were selected by bioinformatics prediction. Five microRNAs, namely miR-371, miR-138, miR-544, miR-145, and miR-214, were confirmed by qRT-PCR and selected as candidate microRNAs. Luciferase reporter assay showed that these five microRNAs could directly target the 3'-UTR of Runx3. However, only simultaneous inhibition of these five microRNAs could alter the expression of Runx3. Most importantly, only simultaneous inhibition could improve the Th1/Th2 balance. Thus, we suggest that miR-371, miR-138, miR-544, miR-145, and miR-214 can modulate the Th1/Th2 balance in asthma by regulating Runx3 in a combinatorial manner.
哮喘与Th1/Th2细胞失衡密切相关，而Runx3在辅助性T细胞分化中起关键作用。本研究旨在调查哮喘患者CD4 T细胞中可能靶向Runx3的失调微小RNA，并揭示Runx3在Th1/Th2平衡调节中的功能。我们通过ELISA检测Th1和Th2相关细胞因子的水平，并通过qRT-PCR分析辅助性T细胞的分化标志物基因。结果表明，我们的哮喘患者存在Th1/Th2细胞失衡。哮喘患者CD4 T细胞中Runx3表达降低。Runx3过表达可恢复Th1/Th2平衡。进行微小RNA微阵列分析后，我们发现哮喘患者CD4 T细胞中有一系列微小RNA发生了显著变化。在这些上调的微小RNA中，通过生物信息学预测选择了8种可能靶向Runx3的微小RNA。通过qRT-PCR确认了5种微小RNA，即miR-371、miR-138、miR-544、miR-145和miR-214，并将其选为候选微小RNA。荧光素酶报告基因检测表明，这5种微小RNA可直接靶向Runx3的3'-UTR。然而，只有同时抑制这5种微小RNA才能改变Runx3的表达。最重要的是，只有同时抑制才能改善Th1/Th2平衡。因此，我们认为miR-371、miR-138、miR-544、miR-145和miR-214可通过组合调节Runx3来调节哮喘中的Th1/Th2平衡。
