Fig. 4

A20 inhibits the invasive and metastatic potential of HCC cells in vitro and in vivo. a-b Cells were plated in 12-well plate and allowed to grow to confluence. Medium was removed and wounds were introduced by scraping the confluent cell cultures with a 200 μL pipette tip. After treatment for 72 h, the wound healing process was monitored under an inverted light microscope. Representative results are shown. Magnification, ×100. Plots (right panel) are mean ± SEM of data from three independent experiments. *, p < 0.05. c-d Effects of A20 depletion (c) or A20 overexpression (d) on the migration and invasion of HCCLM3 cells were assessed by transwell migration assay and matrigel invasion assay, respectively. Representative results are shown in the left panel. Graphs below panels are numbers of migrated cells (mean ± SEM) from three independent experiments. *, p < 0.05. (e-f) LM3-shcon or LM3-shA20 cells (e) or LM3-con or LM3-A20 cells (f) were injected into the tail vein of nude mice (n = 5). Three months post inoculation, mice were sacrificed and metastatic tumor colonies in the lung were examined microscopically. Representative images of H&E staining of lung metastatic foci in each group are shown. The number and the length of metastatic nodules in the lungs of each group is measured and presented as the mean ± SEM below the panels. *, p < 0.05