Fig. 3

Mouse DC-SIGNR knockdown inhibits colon cancer liver metastasis in vivo. a, b The effects of two different mouse DC-SIGNR shRNA constructs on the expression of mouse DC-SIGNR were detected by RT-PCR and Western Blot analysis. Liver tissue from mice injected with a non-targeting shRNA plasmid was used as the positive control. β-actin was used as a loading control. c The effect of different doses of the mouse DC-SIGNR shRNA plasmid on the expression of mouse DC-SIGNR was analysed by RT-PCR. The greyscale analysis of the mouse DC-SIGNR bands was normalized to β-actin. d RT-PCR analysis was used to analyse the mouse DC-SIGNR expression in the mouse livers at the indicated time points following the injection of 60 μg of the mouse DC-SIGNR shRNA plasmid. The greyscale analysis of the mouse DC-SIGNR bands was normalized to β-actin. ***P < 0.001. e, f Liver sections were extracted from mice 3 days after injection with the mDC-SIGNR shRNA plasmid or a control shRNA plasmid via tail vein. LoVo cells that had adhered to frozen liver sections were detected by H&E staining. The adherent colon cancer cells are labelled with red arrows. g Representative sections of liver tumors (n = 5 in each group). The black arrows indicate the tumor nodules. Knocking down mouse DC-SIGNR significantly decreased the metastasis of LoVo and LS174T cells to the liver. h Comparison of the survival percentage of the mouse DC-SIGNR shRNA group and control shRNA group. The survival percentage of mice in the mouse DC-SIGNR shRNA group was significantly higher than that of the control shRNA group for both of the colon cancer cell lines (P = 0.0018 and P = 0.0027, respectively). i Haematoxylin and eosin (H&E)-stained liver and spleen sections are shown. Scale bars, 50 μm and 100 μm, respectively. j The mean volume of tumors and the number of metastases in the liver were determined. k The mean volume of tumors and the number of metastases in the spleen were determined. *P < 0.05, **P < 0.01, ***P < 0.001. The error bars in all graphs represent SD