Fig. 1
AnxA6 overexpression retards cell migration and tumorigenesis. (A) The protein expression of AnxA6 was detected in LO2 and several HCC cell lines by western blotting (up). β-actin was used as an internal control. The quantitative data of AnxA6 expression in LO2 and HCC cells were the means ± SD from three independent experiments (bottom). (B-C) AnxA6 overexpression inhibited cell migration of HepG2 or SK-Hep1. The plasmid pTango-4*Flag-AnxA6 (pFlag-AnxA6) was transiently transfected into HepG2 or SK-Hep1 for 48 h, then cell migration was analyzed by the transwell assay for another 24 h of culture. Images were captured with a microscope, and three random fields were selected to count the number of migration cells. Scale bar represented 20 μm. (D-E) AnxA6 overexpression suppressed mouse tumor growth of the subcutaneous HCC xenograft (D) and orthotopic hepatoma models (E). For subcutaneous HCC xenograft models, 1 × 107 HepG2 or AnxA6-overexpressing HepG2 cells, were subcutaneously injected into each nude mouse with male BALB/c individually, and the tumors in each group (n = 5) were collected to weigh after 4 weeks (D). (E) In the mouse orthotopic HCC model that was injected with wild-type murine Hepa1-6 cells or AnxA6-overexpressing Hepa 1–6 cells, the liver tumors of one mouse were indicated with red dotted line and white arrow (left), and the mean numbers of liver surface tumors respectively from 3 mice were counted (right). ns, no statistical; *P < 0.05; **P < 0.01; ***P < 0.001