Fig. 2

Inhibition of circPDHK1 suppresses the proliferation, migration and invasion of ccRCC cells in vitro and in vivo. A Colony formation assays, (B) CCK8 assays and (C, D) EdU assays were performed to detect the cell viability and proliferation activity of Caki-1 and 786-O cells transfected with two circPDHK1 siRNA. Bars = 200 μm. E–G Representative images and quantification of the transwell migration and invasion assays in circPDHK1 knockdown, and control ccRCC cells. Bars = 100 μm. H-I Representative images and quantification of the transwell migration and invasion assays in circPDHK1 knockdown, and control ccRCC cells. Bars = 100 μm. J Subcutaneous 786-O tumors in NSG mice injected with cholesterol-modified circPDHK1 siRNA, and tumor volumes were calculated every 2 d. At Day 37 after treatment, all mice were sacrificed, and (K) subcutaneous tumors were dissected and recorded. L Subcutaneous tumor was also photographed. (n = 6, each group). M Representative photomicrographs of Ki-67 immunohistochemical (IHC) staining of subcutaneous tumors. N We also established a lung metastasis model by injecting 6 × 10⁶ Caki-1 cells into the tail vein to evaluate the ability of cells to metastasize in vivo. Representative in vivo imaging of mice, photographs of whole lung tissues and hematoxylin–eosin (H&E) staining of lung metastatic nodules. (n = 3, each group) Bars = 200 μm. *P < 0.05; **P < 0.01; ***P < 0.001; ns, no significance