Research Paper Volume 13, Issue 14 pp 18340—18359

CircSPIDR acts as a tumour suppressor in cervical adenocarcinoma by sponging miR-431-5p and regulating SORCS1 and CUBN expression

SORCS1 and CUBN are the functional targets of circSPIDR/miR-431-5p signaling. (A) Western blot analysis of SORCS1 and CUBN protein levels in CADC cells transfected with miR-431-5p inhibitors or the circSPIDR expression vector. (B) AGO2-RIP was performed using an anti-AGO2 antibody in HeLa cells transfected with circSPIDR or the vector. Then, qRT-PCR was used to assess and the enrichment of SORCS1 and CUBN. (C) Western blot analysis of SORCS1 and CUBN protein expression in CADC cells co-transfected with circSPIDR and miR-431-5p mimics. (D) CCK-8 assay in SORCS1- or CUBN-knockdown HeLa cells. (E, F) CCK-8 assay (E) and apoptosis assay (F) in HeLa cells following miR-431-5p inhibition and SORCS1 or CUBN knockdown. (G, H) CCK-8 assay (G) and apoptosis assay (H) in HeLa cells following circSPIDR overexpression and SORCS1 or CUBN knockdown. NS, not significant; *P **P ***P

Figure 7. SORCS1 and CUBN are the functional targets of circSPIDR/miR-431-5p signaling. (A) Western blot analysis of SORCS1 and CUBN protein levels in CADC cells transfected with miR-431-5p inhibitors or the circSPIDR expression vector. (B) AGO2-RIP was performed using an anti-AGO2 antibody in HeLa cells transfected with circSPIDR or the vector. Then, qRT-PCR was used to assess and the enrichment of SORCS1 and CUBN. (C) Western blot analysis of SORCS1 and CUBN protein expression in CADC cells co-transfected with circSPIDR and miR-431-5p mimics. (D) CCK-8 assay in SORCS1- or CUBN-knockdown HeLa cells. (E, F) CCK-8 assay (E) and apoptosis assay (F) in HeLa cells following miR-431-5p inhibition and SORCS1 or CUBN knockdown. (G, H) CCK-8 assay (G) and apoptosis assay (H) in HeLa cells following circSPIDR overexpression and SORCS1 or CUBN knockdown. NS, not significant; *P < 0.05; **P < 0.01; ***P < 0.001.