title: Characterization of genome-wide TFCP2 targets in hepatocellular carcinoma: implication of targets FN1 and TJP1 in metastasis.
abstract: BACKGROUND
  Transcription factor CP2 (TFCP2) is overexpressed in hepatocellular carcinoma(HCC) and correlated with the progression of the disease. Here we report the use of an integrated systems biology approach to identify genome-wide scale map of TFCP2 targets as well as the molecular function and pathways regulated by TFCP2 in HCC.
  
  
  METHODS
  We combined Chromatin immunoprecipitation (ChIP) on chip along with gene expression microarrays to study global transcriptional regulation of TFCP2 in HCC. The biological functions, molecular pathways, and networks associated with TFCP2 were identified using computational approaches. Validation of selected target gene expression and direct binding of TFCP2 to promoters were performed by ChIP -PCR and promoter reporter.
  
  
  RESULTS
  TFCP2 fostered a highly aggressive and metastatic phenotype in different HCC cells. Transcriptome analysis showed that alteration of TFCP2 in HCC cells led to change of genes in biological functions involved in cancer, cellular growth and proliferation, angiogenesis, cell movement and attachment. Pathways related to cell movement and cancer progression were also enriched. A quest for TFCP2-regulated factors contributing to metastasis, by integration of transcriptome and ChIP on chip assay, identified fibronectin 1 (FN1) and tight junction protein 1 (TJP1) as targets of TFCP2, and as key mediators of HCC metastasis. Promoter reporter identified the TFCP2-responsive region, and located the motifs of TFCP2-binding sites in the FN1 promoter, which then was confirmed by ChIP-PCR. We further showed that FN1 inhibition blocks the TFCP2-induced increase in HCC cell aggression, and that overexpression of TFCP2 can rescue the effects of FN1 inhibition. Knock down of TJP1 could also rescue, at least in part, the aggressive effect of TFCP2 knockdown in HCC cells.
  
  
  CONCLUSIONS
  The identification of global targets, molecular pathways and networks associated with TFCP2, together with the discovery of the effect of TFCP2 on FN1 and TJP1 that are involved in metastasis, adds to our understanding of the mechanisms that determine a highly aggressive and metastatic phenotype in hepatocarcinogenesis.
authors.affiliation: Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, 79 QingChun Road, HangZhou, China. zjxu@zju.edu.cn.
authors.forename: Xiao
authors.initials: X
authors.lastname: Xu
authors.affiliation: Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, 79 QingChun Road, HangZhou, China. liuzk@zju.edu.cn.
authors.forename: Zhikun
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Key Lab of Combined Multi-Organ Transplantation, Ministry of Public Health, First Affiliated Hospital, Zhejiang University School of Medicine, HangZhou, China. linzhou19@163.com.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Key Lab of Combined Multi-Organ Transplantation, Ministry of Public Health, First Affiliated Hospital, Zhejiang University School of Medicine, HangZhou, China. xiehy@zju.edu.cn.
authors.forename: Haiyang
authors.initials: H
authors.lastname: Xie
authors.affiliation: Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, 79 QingChun Road, HangZhou, China. chengjun1113@aliyun.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, 79 QingChun Road, HangZhou, China. lingqilynch@163.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Ling
authors.affiliation: Key Lab of Combined Multi-Organ Transplantation, Ministry of Public Health, First Affiliated Hospital, Zhejiang University School of Medicine, HangZhou, China. 21118059@zju.edu.cn.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Wang
authors.affiliation: Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, 79 QingChun Road, HangZhou, China. guohaijun431@126.com.
authors.forename: Haijun
authors.initials: H
authors.lastname: Guo
authors.affiliation: Key Lab of Combined Multi-Organ Transplantation, Ministry of Public Health, First Affiliated Hospital, Zhejiang University School of Medicine, HangZhou, China. weixuyong1986@zju.edu.cn.
authors.forename: Xuyong
authors.initials: X
authors.lastname: Wei
authors.affiliation: Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, 79 QingChun Road, HangZhou, China. zyzss@zju.edu.cn.
authors.forename: Shusen
authors.initials: S
authors.lastname: Zheng
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D047369:Chromatin Immunoprecipitation; D002874:Chromosome Mapping; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D055786:Gene Knockout Techniques; D055106:Genome-Wide Association Study; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D009362:Neoplasm Metastasis; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation; D062826:Zonula Occludens-1 Protein / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; C067625:FN1 protein, human; D005353:Fibronectins; C501906:TFCP2 protein, human; D014157:Transcription Factors; D062826:Zonula Occludens-1 Protein