title: ZIP4, a novel determinant of tumor invasion in hepatocellular carcinoma, contributes to tumor recurrence after liver transplantation.
abstract: OBJECTIVE
  Recently, evidence that Zinc transporter ZRT/IRT-like protein 4 (ZIP4) is involved in invasiveness and apoptosis has emerged in pancreatic cancer and prostate cancer. Our aim was to assess the role of ZIP4 in invasiveness, migration and apoptosis of hepatocellular carcinoma (HCC). The prognostic value of ZIP4 in HCC after liver transplantation was evaluated.
  
  
  METHODS
  The role of ZIP4 in HCC was investigated by overexpressing ZIP4 in BEL7402 and HepG2 cells and inhibiting ZIP4 in HuH-7 and HepG2 cells, using overexpression and shRNA plasmids in vitro studies. Immunohistochemical analysis was used to evaluate ZIP4 expression in HCC tissues from 60 patients undergoing liver transplantation, 36 cirrhotic tissue samples, and 6 normal tissue samples. Prognostic significance was assessed using the Kaplan-Meier method and the log-rank test.
  
  
  RESULTS
  Specific suppression of ZIP4 reduced cell migration and invasiveness, whereas ZIP4 overexpression caused increases in cell migration and invasiveness. Furthermore, overexpression of ZIP4 resulted in increased expression of pro-metastatic genes (MMP-2, MMP-9) and decreased expression of pro-apoptotic genes (caspase-3, caspase-9, Bax). In contrast, suppression of ZIP4 resulted in an opposite effect. ZIP4 was more highly expressed in tumor tissues than non-tumor tissues (P < 0.0001). ZIP4 expression was significantly associated with tumor recurrence (P = 0.002), tumor node metastasis stage (P = 0.044), Child-Turcotte-Pugh score (P = 0.042), and tumor size (P = 0.022). Univariate analysis showed that ZIP4 expression was significantly associated with overall survival (P = 0.020) and tumor-free survival (P = 0.049). Multivariate analysis revealed that ZIP4 was an independent predictor of overall survival (P = 0.037) after liver transplantation.
  
  
  CONCLUSIONS
  ZIP4 could promote migration, invasiveness, and suppress apoptosis in hepatocellular carcinoma, and represent a novel predictor of poor prognosis and therapeutic target for patients with HCC who undergo liver transplantation.
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 3. Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Xu
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Hai-Jun
authors.initials: HJ
authors.lastname: Guo
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Hai-Yang
authors.initials: HY
authors.lastname: Xie
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Jie
authors.initials: J
authors.lastname: Li
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Run-Zhou
authors.initials: RZ
authors.lastname: Zhuang
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Qi
authors.initials: Q
authors.lastname: Ling
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Lin
authors.initials: L
authors.lastname: Zhou
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Xu-Yong
authors.initials: XY
authors.lastname: Wei
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Zhi-Kun
authors.initials: ZK
authors.lastname: Liu
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Song-Ming
authors.initials: SM
authors.lastname: Ding
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Kang-Jie
authors.initials: KJ
authors.lastname: Chen
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 2. Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health and Key Laboratory of Organ Transplantation of Zhejiang Province, Hangzhou, China;
authors.forename: Zhi-Yuan
authors.initials: ZY
authors.lastname: Xu
authors.affiliation: 1. Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated, Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; ; 3. Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China.
authors.forename: Shu-Sen
authors.initials: SS
authors.lastname: Zheng
mesh_terms: D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000401:mortality / Q000473:pathology; D016022:Case-Control Studies; D027682:Cation Transport Proteins / Q000378:metabolism*; D002453:Cell Cycle; D002681:China / Q000453:epidemiology; D005260:Female; D056945:Hep G2 Cells; D006801:Humans; D008099:Liver / Q000473:pathology; D008113:Liver Neoplasms / Q000378:metabolism* / Q000401:mortality / Q000473:pathology; D016031:Liver Transplantation; D008297:Male; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D009364:Neoplasm Recurrence, Local / Q000378:metabolism*
chemical_list: D027682:Cation Transport Proteins; C486791:SLC39A4 protein, human
keywords: ZIP4; hepatocellular carcinoma; liver transplantation