Fig. 3: WFDC21P inhibits HCC metastasis.

a, b Overexpression of WFDC21P mitigates the migratory and invasive capacities of Huh7 cells. Wound healing assays (a), transwell cell migration assays (b, upper), and transwell cell invasion assays (b, lower) of control Huh7 cells and WFDC21P overexpressed Huh7 cells were performed with the pretreatment of mitomycin C (10 μg/mL) for 2 h. The gap sizes of wound healing assay were shown, and the cell migration and invasion numbers of at least five fields were counted. Scale bars in a, b were 500 μm and 200 μm, respectively. c, d knocking down WFDC21P enhances the migratory and invasive capacities of Huh7 cells. e WFDC21P inhibits EMT in HCC cells. WFDC21P was knockdown or overexpressed in Huh7 cells. The protein levels of E-cadherin, N-cadherin, and Vimentin were detected by western blotting. f, g Depletion of WFDC21P promotes tumor metastasis in mice. Luciferase-expressing control and WFDC21P knocking down Huh7 cells were intrasplenically injected into nude mice (n = 6). Liver metastasis was quantified at 58 days after the implantation. Representative bioluminescent images of the mice livers and bioluminescent signal intensities were shown (f). Representative images of livers and liver H&E-staining are shown (g). Scale bars in f, g were 1 cm and 2 mm, respectively. h WFDC21P suppresses EMT in tumor tissues. The expressions of E-cadherin, N-cadherin, and Vimentin were detected in metastatic tumor tissues from g. Scale bars, 100 μm. The data are represented as the means ± SEM of at least three independent experiments. *p < 0.05; and ***p < 0.001.