DLX6-AS1通过竞争性结合miR-15上调Caspase-1诱导巨噬细胞焦亡

DLX6-AS1 Induces Macrophage Pyroptosis Through miR-15/Caspase-1

摘要

为探讨长链非编码RNA (long noncoding RNA, lncRNA) DLX6-AS1在巨噬细胞焦亡中的作用及相关机制, 采用生物信息学分析、实时荧光定量PCR和双荧光素酶报告基因实验等, 在THP-1巨噬细胞中分析并验证lncRNA DLX6-AS1下游微RNA (microRNA, miRNA)及miRNA的下游靶基因; 通过免疫荧光染色等实验检测DLX6-AS1/miR-15/caspase-1轴对巨噬细胞焦亡的影响。结果显示, DLX6-AS1在焦亡巨噬细胞中表达上调, 敲降DLX6-AS1可抑制巨噬细胞焦亡, 但这种抑制作用被miR-15抑制剂逆转; miR-15通过调控其靶基因caspase-1表达, 抑制巨噬细胞焦亡。本研究表明, DLX6-AS1通过竞争性结合miR-15, 解除miR-15对其靶基因caspase-1的抑制作用, 从而诱导巨噬细胞焦亡, 这将丰富lncRNA调控巨噬细胞焦亡的理论基础。

Abstract

In order to investigate the role of long noncoding RNA (lncRNA) DLX6-AS1 in macrophage pyrop-tosis and its related mechanism, the downstream microRNAs (miRNAs) of DLX6-AS1 and the target genes of miRNAs were analyzed in THP-1 cell line by bioinformatics analysis, real-time fluorescent quantitative PCR (qRT-PCR), a dual-luciferase reporter system, and so on. The effects of DLX6-AS1 and downstream miR-15 on macrophage pyroptosis were detected by immunofluorescence staining. The results showed that the expression of DLX6-AS1 was up-regulated in pyroptotic macrophages, and knocking down DLX6-AS1 inhibited pyroptosis of macrophages, which, however, could be reversed by miR-15 inhibitor. Further studies found that miR-15 inhibited macrophage pyroptosis by regulating the target gene caspase-1. These results indicated that DLX6-AS1 induces macrophage pyroptosis by competitively binding to miR-15 and removing the inhibition of miR-15 on its target gene caspase-1. The experiments will enrich the theoretical basis of lncRNA in regulating macrophage pyroptosis.

关键词

长链非编码RNA (lncRNA) / 微RNA (miRNA) / 焦亡 / 巨噬细胞

Key words

long noncoding RNA (lncRNA) / microRNA (miRNA) / pyroptosis / macrophage