LncRNA‐PCAT1 targeting miR‐145‐5p promotes TLR4‐associated osteogenic differentiation of adipose‐derived stem cells

Abstract

This study was aimed to explore the differential expression of long noncoding RNAs (lncRNA)‐PCAT1, miR‐145‐5p and TLR4 in osteogenic differentiation via the Toll‐like receptor (TLR) signalling pathway and consequently determine the potential molecular mechanism. The mRNAs and pathways related to the osteogenic differentiation in human adipose‐derived stem cells (hADSCs) were analysed by bioinformatics. The MiRanda and TargetScan database were employed to detect the potential binding sites of miRNAs on lncRNAs and mRNAs. The differential expression of lncRNA‐PCAT1, miR‐145‐5p and TLR4 were detected by qRT‐PCR. Rrelated protein expression was analysed by Western blot. The targeted relationships between lncRNA‐PCAT1, miR‐145‐5p and TLR4 were verified by dual‐luciferase reporter assay. Alkaline phosphatase (ALP) activity and ARS staining assays were used to measure the impacts exerted by lncRNA PCAT1, miR‐145‐5p and TLR4 mRNA on osteogenic differentiation. After the induction of osteoblast differentiation, the expression of lncRNA‐PCAT1 and TLR4 increased, while the expression of miR‐145‐5p decreased. Dual‐luciferase reporter assay confirmed the targeted relationship between lncRNA‐PCAT1, miR‐145‐5p, and TLR4. LncRNA‐PCAT1 negatively regulated miR‐145‐5p and positively regulated TLR4. Knockdown of lncRNA‐PCAT1 or TLR4 decreased the expression of osteogenic differentiation‐related proteins, reduced the ALP and ARS levels and the activity of the TLR signalling pathway. MiR‐145‐5p could reverse the effects of PCAT1 and TLR4 in hADSCs osteogenic differentiation. LncRNA‐PCAT1 negatively regulated miR‐145‐5p, which promoted TLR4 expression to promote osteogenic differentiation by activating the TLR signalling pathway.

Adipogenesis, Osteogenesis, and Chondrogenesis of Human Mesenchymal Stem/Stromal Cells: A Comparative Transcriptome Approach

“…In addition, another pathway explored was the toll-like receptor (TLR) signaling pathway. Using similar strategies to previous study ( Wu et al, 2018 ), Yu et al (2018) observed an interaction between TLR4 (upregulated in osteogenesis) with lncRNA-PCAT1 and miR-145-5p, which were able to regulate the TLR pathway.…”

Long noncoding RNA LINC00511 induced by SP1 accelerates the glioma progression through targeting miR‐124‐3p/CCND2 axis

“…Emerging studies have demonstrated the increasing vital roles of noncoding RNAs (ncRNAs) in the human diseases, especially cancers . For the glioma, conservatively, hundreds of lncRNAs have been reported to participate in the cellular processes modulation.…”

Long Non-coding RNA HIX003209 Promotes Inflammation by Sponging miR-6089 via TLR4/NF-κB Signaling Pathway in Rheumatoid Arthritis

“…It has been well-documented that many non-coding RNAs can targetedly regulate specific TLRs, and thus contribute to the development of RA, including lncRNAs (15, 16, 30). Many studies have implicated the critical role of lncRNAs in regulating autoimmune and inflammation by targeting TLRs, such as TLR2, TLR4, and TLR3 (35–37). Most interestingly, more and more published studies have suggested that some established lncRNAs can regulate TLR signaling transduction and the relevant immune function in cancer, autoimmune and inflammatory disorders by acting as ceRNAs, such as networks of lncRNA SNHG1-miR-140-TLR4, lncRNA X-miR-154-5p-TLR5, and lncRNA Gm6135-miR-203-3p-TLR4 (38–41).…”