Circ_0045714/miR-331-3p interaction affects IL-1β-evoked human articular chondrocyte injury through regulating PIK3R3 in a ceRNA regulatory cascade

Abstract

Background Osteoarthritis (OA) is characterized by joint pain and joint function limitation. Hsa_circ_0045714 (circ_0045714) is a novel OA-related circular RNA. However, its repertoire remains to be further clarified in joint chondrocytes. Methods RNA and protein expression levels and inflammatory factor levels were detected by real-time quantitative polymerase chain reaction, western blotting and enzyme-linked immunosorbent assay. Cell proliferation and apoptosis were determined by colony formation assay, cell counting kit-8 assay and apoptosis assay. Direct interaction was predicted by bioinformatics method and confirmed by dual-luciferase reporter assay. Results Expression of circ_0045714 and phosphoinositide-3-kinase (PI3K) regulatory subunit 3 (PIK3R3) was declined, and microRNA (miR)-331-3p was promoted in knee articular cartilages and cells from OA patients, as well as interleukin (IL)-1β-challenged human articular chondrocytes (HAC) cell line. In stimulation of IL-1β, HAC cells showed a loss of colony formation ability, cell viability and expression of Bcl-2 and Collagen II, allied with an increase in apoptosis rate and levels of IL-6, IL-8 and tumor necrosis factor-α, Bcl-2-associated X protein, cleaved caspase-3, and ADAM with thrombospondin motif-5. Noticeably, overexpressing circ_0045714 and inhibiting miR-331-3p could suppress IL-1β-evoked these effects, and both were through up-regulating PIK3R3, a key gene in PI3K/AKT signaling pathway. Mechanically, circ_0045714 functioned as competing endogenous RNA (ceRNA) for miR-331-3p and further regulated expression of the downstream target gene PIK3R3. Conclusion There was a novel circ_0045714/miR-331-3p/PIK3R3 ceRNA axis in HAC, and its inhibition might be one mechanism of HAC injury in OA.

Circ_0005526 contributes to interleukin-1β-induced chondrocyte injury in osteoarthritis via upregulating transcription factor 4 by interacting with miR-142-5p

“… CircRNAs are identified to be associated with the pathogenesis of OA [ 22 ]. Hsa_circ_0045714 level downregulation reduced IL-1β-evoked OA chondrocyte injury [ 23 ]. Silencing circ_0001846 alleviated cell apoptosis and inflammation in IL-1β-induced OA progression [ 24 ].…”

“…Additionally, circ_0134111 promoted OA chondrocyte damage by inhibiting miR-224-5p to induce overexpression of CC motif ligand 1 (CCL1) [ 33 ]. Circ_0045714 increased phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1) level to regulate IL-1β-stimulated chondrocyte injury via serving as a ceRNA of miR-331-3p [ 23 ]. In this study, miR-142-5p directly targeted TCF4 and circ_0005526 led to upregulation of TCF4 expression via targeting miR-142-5p.…”

The therapeutic effect and mechanism of melatonin on osteoarthritis: From the perspective of non-coding RNAs

“… Li et al (2017) found that circ 0045714 could regulate the synthesis of ECM by inhibiting the transcriptional activity of miR-193b and promoting the expression of miR-193b target gene insulin-like growth factor 1 receptor (IGF1R) ( Li et al, 2017 ). Ding et al (2021) .up found that circ 0045714 plays a protective role in IL-1β-induced chondrocyte injury by targeting and inhibiting miR-331-3p and then upregulating PIK3R3, which is manifested in alleviating chondrocyte apoptosis, inflammatory response and ECM degradation ( Ding et al, 2021 ).…”

Circ_0044235 regulates the development of osteoarthritis by the modulation of miR-375/PIK3R3 axis

“…Phosphoinositide 3-kinase (PI3K) regulatory subunit 3 (PIK3R3) is an inhibitor of PI3K [ 19 ] in the PI3K/AKT pathway, and is intimately related to OA pathogenesis [ 20 , 21 ]. PIK3R3 was confirmed to be downregulated in OA knee cartilage and participated in OA progression [ 22 , 23 ]. Here, we showed that circ_0044235 could bind to miR-375, and miR-375 could target PIK3R3.…”