LncRNA NEAT1参与TLR2介导的炎症因子的表达

Identification LncRNA NEAT1 as a new mediator of the TLR2-induced cytokine production

长链非编码RNA(long non-coding RNA,lncRNA)被发现在真核生物体内广泛表达,并且可以参与表观调节、基因转录、转录后调节等生物学过程。本研究旨在探索lncRNA NEAT1在先天免疫反应中的作用,尤其是对TLR2通路的调节。应用RT-PCR及核糖核酸酶保护实验的方法检测lncRNA NEAT1受TLR2刺激剂Pam3CKS4的诱导情况。利用反义寡核苷酸(antisense oligonucleotides,ASO)转染THP-1用以沉默NEAT1的表达,并进一步研究其功能。结果显示Pam3CKS4可以诱导NEAT1的表达,尤其是NEAT1-V1。并且沉默NEAT1的表达能明显下调一部分TLR2信号活化诱导的细胞因子、趋化因子的表达,包括IL6、CXCL10、CCL2、CCL8和CXCL11等,但是NEAT1对TLR2信号活化诱导的TNF-α、IL-1β的表达没有明显影响。同时我们发现受NEAT1调节的基因均为TLR2持续缓慢诱导基因,而TLR2信号的早期反应基因TNF-α、IL-1β不受NEAT1调节。以上结果说明lncRNA NEAT1参与TLR2介导的先天免疫调节,并且选择性地调控一组TLR2信号活化缓慢持续诱导的晚期反应炎症因子的表达。

Long noncoding RNAs （lncRNAs） have recently been identified to be expressed in eukaryotes, and tightly linked to diverse biological processes through affecting epigenetic regulation, gene transcription and post-transcription regulation and so on. This study was undertaken to investigate whether IncRNA NEAT1 would be involved in innate immune response, in special the contribution of the IncRNA NEAT1 in the TLR2 signaling pathway. RT-PCR and ribonuclease protection assay（RPA） were used to detect NEAT1 expression after Pam3CKS4 stimulation. Transfection of antisense oligonucleotides （ASO） was conducted to silence NEAT1 and determine the biologic function of NEAT1. The results were shown that NEAT1 expression was abnormally increased after Pam3CKS4 stimulation. Furthermore we found silencing NEAT1 significantly reduced the ex- pression of a group of cytokines, including IL-6, CXCL10, CCL2, CCL8 and CXCLll etc. , rather than TNF-αand IL-1β. Im- portantly, we noticed that the NEATl-regulated genes were induced by Pam3CKS4 continuously and in late stages. In conclu- sions, NEAT1 could participate in the regulation of the TLR2 signaling pathway, and selectively regulated a subset of Pam3CKS4-induced inflammatory factors in late stages.