a The Rhinology Department , The First Affiliated Hospital of Zhengzhou University , Zhengzhou , China.
b The Academy of medical sciences , Zhengzhou University , Zhengzhou , China.
Cell Cycle. 2019 Feb;18(3):312-319. doi: 10.1080/15384101.2018.1562285. Epub 2019 Jan 17.
The aim of this study was to investigate the role of lncRNA NEAT1 (nuclear enriched abundant transcript 1) in regulating Th2 cell differentiation. The overexpression vectors of NEAT1 and ITCH, and siRNA targeting NEAT1, EZH2 (enhancer of zeste homolog 2) and STAT6 (signal transducer and activator of transcription 6) were transfected into CD4+T cells. The mRNA expressions of ITCH and STAT6 were analyzed by qRT-PCR and western blotting. The levels of Th2 cytokines were detected by ELISA assay. RIP and ChIP assays were performed to analyze the association between NEAT1 and EZH2 as well as EZH2 and STAT6, respectively. Results showed that NEAT1 significantly repressed ITCH expression and increased STAT6 expression as well as the levels of IL-4, IL-5 and IL-13 in CD4+T cells. RIP and ChIP assays revealed that NEAT1 bound to EZH2 and EZH2 was recruited to the promoter region of ITCH in CD4+T cells. Silencing EZH2 significantly promoted STAT6 for ubiquitination. Furthermore, NEAT targeted STAT6 for ubiquitination and elevated levels of Th2 cytokines by regulating EZH2/ITCH axis. In conclusion, our data indicated that NEAT1 promotes Th2 cell differentiation through the EZH2/ITCH/STAT6 axis.
本研究旨在探讨长链非编码 RNA NEAT1（核丰富丰富转录物 1）在调节 Th2 细胞分化中的作用。将 NEAT1 和 ITCH 的过表达载体以及针对 NEAT1、EZH2（增强子结合锌指蛋白 2）和 STAT6（信号转导和转录激活因子 6）的 siRNA 转染到 CD4+T 细胞中。通过 qRT-PCR 和 Western blot 分析 ITCH 和 STAT6 的 mRNA 表达。通过 ELISA 测定法检测 Th2 细胞因子的水平。RIP 和 ChIP 测定分别分析了 NEAT1 与 EZH2 以及 EZH2 与 STAT6 之间的关联。结果表明，NEAT1 可显著抑制 ITCH 的表达并增加 STAT6 的表达以及 CD4+T 细胞中 IL-4、IL-5 和 IL-13 的水平。RIP 和 ChIP 测定表明，NEAT1 与 EZH2 结合，EZH2 被招募到 CD4+T 细胞中 ITCH 的启动子区域。沉默 EZH2 可显著促进 STAT6 的泛素化。此外，NEAT1 通过调节 EZH2/ITCH 轴促进 STAT6 的泛素化和 Th2 细胞因子的水平升高。总之，我们的数据表明，NEAT1 通过 EZH2/ITCH/STAT6 轴促进 Th2 细胞分化。
