Fig. 3
Knockdown of circPDE5A promotes ESCC proliferation and metastasis in vitro and in vivo. A RT-qPCR detection of linear PDE5A transcript and circPDE5A level in ECA109 cells after treatment with sh-NC, sh-circPDE5A-1, sh-circPDE5A-2, and sh-circPDE5A-3 plasmid. B–D The proliferative capacity of ECA109 cells assessed by CCK8 (B), plate cloning (C), and EdU (D) assays after treatment with sh-NC, sh-circPDE5A-1, and sh-circPDE5A-2 plasmid. Bar in (D) represents 50 μm. E Motility of ECA109 cells determined by wound healing assay after treatment with sh-NC, sh-circPDE5A-1, and sh-circPDE5A-2 plasmid. Bar represents 100 μm. F Migration and invasion ability of ECA109 cells determined by transwell assay after treatment with sh-NC, sh-circPDE5A-1, and sh-circPDE5A-2 plasmid. Bar represents 1,000 μm. G EMT marker protein levels in ECA109 cells detected by WB after treatment with sh-NC, sh-circPDE5A-1, and sh-circPDE5A-2 plasmid. H Images of xenograft tumors formed in nude mice by subcutaneous injection of ECA109 cells transfected with sh-NC, sh-circPDE5A-1, and sh-circPDE5A-2, respectively. I Volume growth curves of nude mouse xenograft tumors. Tumor volume measured every 7 days after inoculation for 6 weeks (n = 6, unit: mm3). J The weight of xenograft tumors in nude mice measured at the end of the experiment (n = 6, unit: mg). K Representative bioluminescent images of lung metastases formed by tail vein injection of ECA109 cells transfected with sh-NC, sh-circPDE5A-1, and sh-circPDE5A-2, respectively (n = 6). L Diagram and HE staining of representative lung tissue from each group. Bar represents 2,000 μm. M Number of metastatic lung nodules in each group (n = 6). ***P < 0.005. ns, not significant