Regulatory mechanism of hsa_circ_0072697 in AS-PBMCs
To investigate the regulatory mechanism of hsa_circ_0072697 in PBMCs of AS patients, we performed overexpression and knockdown of hsa_circ_0072697 in PBMCs, and divided them into 5 groups: AS-PBMC, AS-PBMC+pcDNA3.1 hsa_circ_0072697-NC, AS-PBMC+pcDNA3.1 hsa_circ_0072697, AS-PBMC+siRNA-hsa_circ_0072697-NC, AS-PBMC+siRNA-hsa_circ_0072697. CCK-8 assay showed that the cell proliferation activity of the hsa_circ_0072697 overexpression group was significantly reduced compared to the other four groups, while the cell proliferation activity of the siRNA-hsa_circ_0072697 group was increased (Fig. 9A). These results suggest that hsa_circ_0072697 may play an inhibitory role in the pathogenesis of AS. Furthermore, apoptosis analysis by flow cytometry showed that the hsa_circ_0072697 overexpression group had the highest apoptosis rate, while the siRNA-hsa_circ_0072697 group had the lowest apoptosis rate (Fig. 9B). This indicates that hsa_circ_0072697 is also involved in regulating the apoptosis of PBMCs. Finally, from cell cycle analysis performed by flow cytometry, we observed that the hsa_circ_0072697 overexpression group had a decrease in the number of cells in the G1 phase and an increase in the number of cells in the S and G2 phases compared to the control group, while the siRNA-hsa_circ_0072697 group had completely opposite effects on the cell cycle of PBMCs (Fig. 9C). These findings indicate that hsa_circ_0072697 plays an important role in the cell cycle of PBMCs.