Fig. 7.

Knockdown of supervillin inhibits HCC metastasis. Mice (n = 5) were implanted with a ~ 1 mm³ tumor “seed” derived from a supervillin-knockdown clone or a clone treated with a control shRNA; mice were treated with or without hepatic artery ligation (HAL). a After 6 weeks, each tumor was dissected, lysed, and the level of supervillin was determined by immunoblotting. β-actin was used as the loading control. Data represent the mean of at least three independent experiments ± SD in MHCC-97H cells. b IHC staining showing the accumulation of HIF1α in mice with HAL, compared with those without HAL. Scale bar = 100 μm. c Bioluminescence imaging of representative mice at the end of the experiment. d Bioluminescence signal intensity in dissected livers was compared among the four experimental groups. e Hematoxylin/eosin staining of lung tissues. Arrows indicate the metastatic tumor foci in the lung tissues. Scale bar = 100 μm. f The bioluminescent signal intensities in the lungs were compared among the four experimental groups. g A schematic diagram showing the signaling axis of supervillin-RhoA/ROCK-ERK/p38 in HCC. Under hypoxia, supervillin levels increase and lead to RhoA activation. Activated RhoA regulates actin polymerization, induces the ROCK-ERK/p38 signaling pathway, and EMT, thereby promoting HCC cell migration, invasiveness, and metastasis