Regulatory effects of hsa_circ_0072697 on AS-related factors
in AS-PBMCs
We performed overexpression and knockdown of hsa_circ_0072697 in
AS-PBMCs and evaluated the relative mRNA expression levels of
hsa-miR-145-5p, MMP3, MMP9, IKB-α, and NF-κB p65 using RT-qPCR
(Fig.11A). Our results demonstrated that the relative mRNA expression
levels of hsa-miR-145-5p, MMP3, MMP9, and NF-κB p65 were downregulated
in AS-PBMC+pcDNA3.1 hsa_circ_0072697 group and upregulated in
AS-PBMC+siRNA-hsa_circ_0072697 group. Conversely, the expression of
NF-κB p65 inhibitor IKB-α was upregulated in AS-PBMC+pcDNA3.1
hsa_circ_0072697 group and downregulated in
AS-PBMC+siRNA-hsa_circ_0072697 group. Furthermore, WB assay showed
that the protein levels of MMP3, MMP9, and p-NF-κB/NF-κB were reduced in
AS-PBMC+pcDNA3.1 hsa_circ_0072697 group and increased in
AS-PBMC+siRNA-hsa_circ_0072697 group. Meanwhile, the protein
expression level of IKBα/p-IKBα was upregulated in AS-PBMC+pcDNA3.1
hsa_circ_0072697 group and downregulated in
AS-PBMC+siRNA-hsa_circ_0072697 group (Fig.11B and 11C). These results
indicated that the expression of hsa_circ_0072697 positively regulates
IKBα/p-IKBα, and negatively regulates MMP3, MMP9, p-NF-κB/NF-κB.