Fig. 6
IL27RA promoted liver cancer progression in a TGFβR1 signalling-dependent manner. Huh-7 and SNU398 cells stably transfected with IL27RA overexpression and knockdown of TGFβR1 were used for immunoblotting analysis. (B) The effect of silencing TGFβR1 on the proliferation of HCC cells overexpressing IL27RA was measured via the CCK8 assay. (C, D) Colony formation assay: The left panel shows representative images, and the right panel presents the quantification analysis. (E–G) Hepa 1‒6 cells with stable overexpression of IL27RA and knockdown of TGFβR1 were injected through the hepatic portal vein into C57BL/6 mice. Representative liver images are shown, and the liver/body weight ratio was determined (right panel). H&E staining revealed liver cancer nodules (n = 10). (F) The ratio of liver weight to total body weight was determined (n = 4). (G) Kaplan‒Meier survival analysis was performed to evaluate the overall survival of the mice (n = 6). (H–I) Transwell migration assays were performed to assess the migration ability of HCC cells overexpressing IL27RA following the silencing of TGFβR1. (J–M) Wound healing analysis was performed to assess the migration of HCC cells at 0 and 48 h. Representative images (left panel) and quantification (right panel) are shown as indicated. (N) Hepa 1‒6 cells with stable overexpression of IL27RA and knockdown of TGFβR1 were injected into the spleens of C57BL/6 mice. After 30 days, the livers of the mice were excised and photographed, and a representative liver is shown. H&E staining revealed liver metastatic nodules (n = 10). (O) The number of liver metastatic nodules was determined and analysed via Student’s t- test (n = 4). (P) Kaplan‒Meier survival analysis was performed to evaluate the OS of the mice (n = 6). All human HCC cells were treated with rmIL-27 (100 ng/ml) for 24 h. The data are presented as the mean ± SDs. **p < 0.01; ***p < 0.001; ****p < 0.0001.
