Abnormal hsa_circ_0003948 expression affects chronic periodontitis development by regulating miR‐144‐3p/NR2F2/PTEN signaling

Abstract

Background and Objective This study aimed to investigate the correlation between chronic periodontitis (CP) and abnormal circular RNA (circRNA) expression and to identify the role of hsa_circ_0003948 in the progression of CP. Methods Next‐generation sequencing was utilized to investigate abnormal expression of circRNA in gingival tissues from CP patients and healthy control subjects. Bioinformatics and luciferase reporting analyses were used to clarify the interactive relationship among circRNA, miRNA, and mRNA. Periodontal ligament cells (PDLCs) were employed to analyze proliferation and apoptosis after lipopolysaccharide (LPS) treatment using the cell counting kit 8 (CCK8) assay and flow cytometry detection. Results High‐throughput sequencing and RT‐qPCR analyses confirmed that hsa_circ_0003948 expression decreased dramatically in gingival samples of CP patients. Overexpression of hsa_circ_0003948 alleviated LPS‐induced PDLC injury by regulating NR2F2/PTEN signaling. The miR‐144‐3p and NR2F2 were determined to be hsa_circ_0003948 downstream targets. NR2F2 downregulation or miR‐144‐3p overexpression reversed the protective effect of hsa_circ_0003948 in PDLCs after treatment with LPS. Upregulation of NR2F2 reversed the inhibitory effect of miR‐144‐3p on surviving PDLCs after LPS treatment. Conclusion Overexpression of hsa_circ_0003948 exerts a protective effect in CP via miR‐144‐3p/NR2F2/PTEN signaling regulation.

Identification of potentially functional circRNAs and prediction of circRNA‐miRNA‐mRNA regulatory network in periodontitis: Bridging the gap between bioinformatics and clinical needs

“…Among these circRNAs, further validation studies suggested that circSERPINE1 has a negative impact on inflammatory response and oxidative stress by regulating the miR‐378h /RORA axis, while circCTHRC1 suppressed cytokine production via sponging let‐7f‐5p to regulate SOCS6 19,21 . CircMAP3K11 promoted the proliferation and inhibited the apoptosis of hPDLSCs via the miR‐511‐3p/TLR4 axis, and circKDELR2 promoted cell proliferation and inhibited apoptosis in response to LPS via the miR‐144‐3p/NR2F2/PTEN axis 18,23 …”

“…51 Several circRNAs have been suggested that might exert regulations in hPDLSCs cell viability, proliferation, migration, apoptosis, osteogenic differentiation, inflammatory response, oxidative stress, cytokine production, as well as stemness maintenance, including circSERPINE1, circCTHRC1, circMAP3K11, circKDELR2, circCDK8, and CDR1as. 14,15,[17][18][19][20][21]23 Among these circRNAs, further validation studies suggested that circSERPINE1 has a negative impact on inflammatory response and oxidative stress by regulating the miR-378h /RORA axis, while circCTHRC1 suppressed cytokine production via sponging let-7f-5p to regulate SOCS6. 19,21 CircMAP3K11…”

“…promoted the proliferation and inhibited the apoptosis of hPDLSCs via the miR-511-3p/TLR4 axis, and circKDELR2 promoted cell proliferation and inhibited apoptosis in response to LPS via the miR-144-3p/NR2F2/PTEN axis. 18,23 For human gingival fibroblasts, circ-Amotl1 could promote cell adhesion, migration, proliferation, and wound healing through suppressing miR-17-5p and increasing the expression of fibronectin, Dnmt3a, and Stat3. 24 On the other hand, pyroptosis of gingival fibroblasts could be induced by circ_0138959, which was accompanied by increased levels of pro-inflammatory cytokines and pyroptosisrelated proteins.…”

“…In addition to the regulation of osteogenic differentiation, other functional validation studies suggested that circRNAs are also involved in the regulation of hPDLSCs cell viability, apoptosis, proliferation, migration, response to inflammation and oxidative stress, and the maintenance of stemness 17–21 . Some recent studies have used human gingival tissue samples to profile circRNA expression, including studies that profiled circRNA expression in human gingival tissues from patients with periodontitis and loss‐of‐function studies that presented circRNA as a promising therapeutical target for periodontitis using human gingival fibroblasts derived from human gingival tissues 22–25 …”

“…[17][18][19][20][21] Some recent studies have used human gingival tissue samples to profile circRNA expression, including studies that profiled circRNA expression in human gingival tissues from patients with periodontitis and loss-of-function studies that presented circRNA as a promising therapeutical target for periodontitis using human gingival fibroblasts derived from human gingival tissues. [22][23][24][25] While studies on the regulation of circRNA in periodontitis are progressing, emerging roles of circRNAs, such as RNA-binding protein sponges, regulators of splicing and transcription, translation, and biomarkers, could act as a "game changer." 7,8,10,[26][27][28] Fortunately, multiple bioinformatic approaches have been developed to help us cope with challenges for circRNA detection and functional prediction.…”