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.