RhoC/ ROCK2 promotes vasculogenic mimicry formation primarily through ERK/MMPs in hepatocellular carcinoma

Abstract

Introduction: Our previous study reported that rho-associated coiled-coil containing kinases (ROCKs) were involved in vasculogenic mimicry (VM) formation in hepatocellular carcinoma (HCC). This study focused on clarifying potential mechanisms of RhoC/ROCK on VM formation in hepatocellular carcinoma (HCC). Method: In total, 80 cases of HCC tissues and adjacent non-tumorous liver tissues were used to identify VM in HCC tissues by PAS/CD34 double staining. Kaplan-Meier analysis was conducted to test the predictive relationship between RhoC/ROCK expression and patient prognosis. Furthermore, we stably overexpressed RhoC and inhibited RhoC, ROCK1 or ROCK2 expression to clarify the effect of RhoC/ROCK on VM formation in vitro and in vivo. Result: RhoC expression was up regulated in HCC tissues, especially the VM-positive (VM+) group, compared to non-cancerous tissues (P < 0.01), and patients with high expression of RhoC had shorter survival times (P < 0.001). Cell motility and VM formation significantly decreased after knockdown of RhoC, while overexpressing RhoC led to the opposite result. Compared to ROCK1 shRNA, ROCK2 shRNA could largely affects VM formation, cell motility and the key VM factors and the epithelial-mesenchymal transition (EMT) markers in vitro and in vivo. Moreover, p-ERK, p-MEK, p-FAK and p-paxillin levels clearly altered following the change of RhoC, but ROCK2 little affected expression of p-FAK. Conclusion: RhoC/ROCK2 may have a major effect on VM in HCC via ERK/MMPs signaling and might be a potential therapeutic target for the treatment of HCC.