Zhihong Yang1, Baocun Sun2, Yanlei Li3, Xiulan Zhao4, Xueming Zhao5, Qiang Gu6, Jindan An7, Xueyi Dong8, Fang Liu9, Yong Wang10. 1. Department of Pathology, Tianjin Medical University, Tianjin 300070, China; Department of Pathology, General Hospital of Tianjin Medical University, Tianjin 300052, China. Electronic address: yangzhihong321@sina.com. 2. Department of Pathology, Tianjin Medical University, Tianjin 300070, China; Department of Pathology, Cancer Hospital of Tianjin Medical University, Tianjin 300060, China; Department of Pathology, General Hospital of Tianjin Medical University, Tianjin 300052, China. Electronic address: baocunsun@gmail.com. 3. Department of Pathology, Tianjin Medical University, Tianjin 300070, China. Electronic address: dylylnj@163.com. 4. Department of Pathology, Tianjin Medical University, Tianjin 300070, China; Department of Pathology, General Hospital of Tianjin Medical University, Tianjin 300052, China. Electronic address: xiulanzhao01@163.com. 5. Department of Pathology, Tianjin Medical University, Tianjin 300070, China. Electronic address: xuemingzhao@163.com. 6. Department of Pathology, Tianjin Medical University, Tianjin 300070, China; Department of Pathology, General Hospital of Tianjin Medical University, Tianjin 300052, China. Electronic address: qiang00gu@163.com. 7. Department of Pathology, Tianjin Medical University, Tianjin 300070, China. Electronic address: jindananan@163.com. 8. Department of Pathology, Tianjin Medical University, Tianjin 300070, China. Electronic address: xueyidongdong@126.com. 9. Department of Pathology, Tianjin Medical University, Tianjin 300070, China. Electronic address: fangliu112233@163.com. 10. Department of Pathology, Tianjin Medical University, Tianjin 300070, China. Electronic address: yongwangwywy@163.com.
Abstract
AIMS: Zinc finger E-box binding homeobox 2 (ZEB2), an epithelial-mesenchymal transition (EMT) regulator, has been involved in invasion and metastasis of human tumor. Although EMT may be involved in vasculogenic mimicry (VM) formation, no reports describing the relation between ZEB2 and VM are available. We hypothesize that ZEB2 may promote VM formation in hepatocellular carcinoma (HCC). METHODS AND RESULTS: Paraffin-embedded tumor tissue samples from 92 patients were immunostained with anti-ZEB2 antibody. We found that the ZEB2 nuclear expression was significantly associated with VM formation and metastasis. Patients with VM and ZEB2 nuclear expression had a shorter survival period than those without expression. In vitro, ZEB2 overexpression significantly enhanced cell motility, invasiveness, and VM formation of HepG2 cells. ZEB2 upregulation also increased VE-cadherin, Flt-1, and Flk-1 expression and activated MMPs. ZEB2 knockdown inhibited cell motility, invasiveness, and VM formation in Bel7402 cells. ZEB2 knockdown also decreased VE-cadherin, Flt-1, and Flk-1 expression and MMP activity. In addition, EMT in HepG2 cells was induced by TGF-β1 treatment, and the kinetics of expression of EMT markers and regulators were assessed by Western blot analysis. The expression of ZEB2 increased significantly, and VM formation was promoted. CONCLUSION: ZEB2 can promote VM formation through the EMT pathway. Our findings may represent a novel therapeutic target in HCC.
AIMS: Zinc finger E-box binding homeobox 2 (ZEB2), an epithelial-mesenchymal transition (EMT) regulator, has been involved in invasion and metastasis of humantumor. Although EMT may be involved in vasculogenic mimicry (VM) formation, no reports describing the relation between ZEB2 and VM are available. We hypothesize that ZEB2 may promote VM formation in hepatocellular carcinoma (HCC). METHODS AND RESULTS:Paraffin-embedded tumor tissue samples from 92 patients were immunostained with anti-ZEB2 antibody. We found that the ZEB2 nuclear expression was significantly associated with VM formation and metastasis. Patients with VM and ZEB2 nuclear expression had a shorter survival period than those without expression. In vitro, ZEB2 overexpression significantly enhanced cell motility, invasiveness, and VM formation of HepG2 cells. ZEB2 upregulation also increased VE-cadherin, Flt-1, and Flk-1 expression and activated MMPs. ZEB2 knockdown inhibited cell motility, invasiveness, and VM formation in Bel7402 cells. ZEB2 knockdown also decreased VE-cadherin, Flt-1, and Flk-1 expression and MMP activity. In addition, EMT in HepG2 cells was induced by TGF-β1 treatment, and the kinetics of expression of EMT markers and regulators were assessed by Western blot analysis. The expression of ZEB2 increased significantly, and VM formation was promoted. CONCLUSION:ZEB2 can promote VM formation through the EMT pathway. Our findings may represent a novel therapeutic target in HCC.