Fig. 3: Sec62 regulates cellular movement by targeting integrin α signalling.

a Left: Functional pathway analysis of the differentially expressed genes was conducted using commercially available IPA software. The change in integrin signalling in both Huh7-Sec62-shRNA cells and Huh7-Sec62-overexpressing cells is consistent with the expression of Sec62. The colour indicates the degree of down- (blue) or up- (orange) regulation following Sec62 knockdown or overexpression in Huh7 cells. Upper right: Cellular movement regulated by Sec62 was identified in both Huh7-Sec62-RNAi cells and Huh7-Sec62-overexpressing cells by GO functional analysis. Lower right: Interaction of Sec62 and integrin signalling after Sec 62 knockdown in Huh7 cells, which was determined based on the integrin signalling interactome in the Ingenuity IPA database overlaid with microarray data from Huh7-Sec62-RNAi cells. b Left: Integrin α5 and integrin αV were overexpressed in Sec62-knockdown (Sec62⁻) Huh 7 cells, and the healing ability of these cells was evaluated by a monolayer wounding assay. Western blot analysis of integrin α 5 and integrin αV expression in Sec62-knockdown cells. Middle: Transwell assay analysis of the migration ability of Sec62⁻/ integrin α5 (top) or Sec62⁻/ integrin αV (low) cells. The number of cells that invaded through the filter into the lower compartment was determined using a colorimetric crystal violet assay. Data are presented as the means ± SD of at least three independent experiments and were compared to the amounts of invaded cells from negative control transfected cells. *P < 0.01 versus the control group (Student’s t test). Right: Western blot of co-immunoprecipitated integrin αV (top) and integrin α5 (low) in Huh7 cells after Co-IP with an anti-Sec62 antibody. c The levels of integrin αV, integrin α 5, CAV1, calpains and MLCK expression in the integrin α/CAV1 pathway from HCC patient samples with recurrence (n = 4) or without recurrence (n = 4) were evaluated by Western blot analysis.