LncRNA HOTAIR regulates the PI3K/AKT pathway via the miR-126-3p/PIK3R2
axis to participate in synovial angiogenesis in rheumatoid arthritis.
Abstract
Abstract: Background: The abnormal expression of lncRNA HOTAIR has been
associated with synovial angiogenesis in rheumatoid arthritis (RA). The
aim of this study is to investigate whether lncRNA HOTAIR can
participate in synovial angiogenesis in RA by regulating the PI3K/AKT
pathway through the miR-126/PIK3R2 axis. Methods: In this study, we
conducted in vitro experiments by designing overexpression plasmids and
small interfering RNAs targeting lncRNA HOTAIR and then transfected them
into fibroblast-like synoviocytes (FLS) obtained from RA patients. We
then co-cultured the modified FLS with human umbilical vein endothelial
cells (HUVEC) to establish an RA-FLS-induced HUVEC model. We
investigated the effects of lncRNA HOTAIR on the proliferation,
migration, and tube formation abilities of HUVECs, as well as the
expression of synovial endothelial cell markers, angiogenic factors, and
the PI3K/AKT pathway. To validate the interactions between lncRNA
HOTAIR, miR-126-3p, and PIK3R2, we used bioinformatics and luciferase
reporter experiments. We also used a combination of real-time
fluorescence quantitative (RT-qPCR), Western blotting (WB), and
immunofluorescence (IF) methods to identify target genes and proteins.
Results: LncRNA HOTAIR was highly expressed in HUVEC cells induced by
RA-FLS. Overexpression of lncRNA HOTAIR significantly increased the
expression of VEGF, bFGF, CD34 and CD105 in HUVEC cells, promoted their
proliferation, invasion, and tube formation, while the silencing of
lncRNA HOTAIR reversed these effects, and the PI3K/AKT activator also
reversed them. Overexpression of lncRNA HOTAIR activated the PI3K/AKT
pathway, promoting the expression of PI3K, AKT, and P-AKT proteins.
Bioinformatics and dual-luciferase assays verified the targeting
relationship between LncRNA HOTAIR, miR-126-3p, and PIK3R2. Conclusion:
LncRNA HOTAIR can activate the PI3K/AKT pathway, possibly through the
regulatory axis of miR-126-3p/PIK3R2, and thus participate in synovial
angiogenesis in rheumatoid arthritis.