title: The transcription factor SALL4 regulates stemness of EpCAM-positive hepatocellular carcinoma.
abstract: OBJECTIVE
  Recent evidence suggests that hepatocellular carcinoma can be classified into certain molecular subtypes with distinct prognoses based on the stem/maturational status of the tumor. We investigated the transcription program deregulated in hepatocellular carcinomas with stem cell features.
  
  
  METHODS
  Gene and protein expression profiles were obtained from 238 (analyzed by microarray), 144 (analyzed by immunohistochemistry), and 61 (analyzed by qRT-PCR) hepatocellular carcinoma cases. Activation/suppression of an identified transcription factor was used to evaluate its role in cell lines. The relationship of the transcription factor and prognosis was statistically examined.
  
  
  RESULTS
  The transcription factor SALL4, known to regulate stemness in embryonic and hematopoietic stem cells, was found to be activated in a hepatocellular carcinoma subtype with stem cell features. SALL4-positive hepatocellular carcinoma patients were associated with high values of serum alpha fetoprotein, high frequency of hepatitis B virus infection, and poor prognosis after surgery compared with SALL4-negative patients. Activation of SALL4 enhanced spheroid formation and invasion capacities, key characteristics of cancer stem cells, and up-regulated the hepatic stem cell markers KRT19, EPCAM, and CD44 in cell lines. Knockdown of SALL4 resulted in the down-regulation of these stem cell markers, together with attenuation of the invasion capacity. The SALL4 expression status was associated with histone deacetylase activity in cell lines, and the histone deacetylase inhibitor successfully suppressed proliferation of SALL4-positive hepatocellular carcinoma cells.
  
  
  CONCLUSIONS
  SALL4 is a valuable biomarker and therapeutic target for the diagnosis and treatment of hepatocellular carcinoma with stem cell features.
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Sha Sha
authors.initials: SS
authors.lastname: Zeng
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan; Department of General Medicine, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan. Electronic address: taroy@m-kanazawa.jp.
authors.forename: Taro
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Mitsumasa
authors.initials: M
authors.lastname: Kondo
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Kouki
authors.initials: K
authors.lastname: Nio
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Hayashi
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Hara
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Yoshimoto
authors.initials: Y
authors.lastname: Nomura
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Mariko
authors.initials: M
authors.lastname: Yoshida
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Hayashi
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Oishi
authors.affiliation: Department of Pathology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Ikeda
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Masao
authors.initials: M
authors.lastname: Honda
authors.affiliation: Department of Gastroenterology, Kanazawa University Hospital, Kanazawa, Ishikawa, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Kaneko
mesh_terms: D000368:Aged; D000951:Antigens, Neoplasm / Q000032:analysis*; D006528:Carcinoma, Hepatocellular / Q000737:chemistry / Q000473:pathology*; D015815:Cell Adhesion Molecules / Q000032:analysis*; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D006655:Histone Deacetylases / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D008113:Liver Neoplasms / Q000737:chemistry / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D014411:Neoplastic Stem Cells / Q000737:chemistry*; D014157:Transcription Factors / Q000032:analysis / Q000502:physiology*; D000509:alpha-Fetoproteins
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; C472575:SALL4 protein, human; D014157:Transcription Factors; D000509:alpha-Fetoproteins; D006655:Histone Deacetylases
keywords: AFP; CSC; Cancer stem cell; Chemosensitivity; EpCAM; Gene expression profile; HCC; HDAC; Hepatocellular carcinoma; HpSC-HCC; MH-HCC; NuRD; SAHA; SALL4; SBHA; Sal-like 4 (Drosophila); alpha fetoprotein; cancer stem cell; epithelial cell adhesion molecule; hepatic stem cell-like HCC; hepatocellular carcinoma; histone deacetylase; mature hepatocyte-like HCC; nucleosome remodeling and deacetylase; qRT-PCR; quantitative reverse transcription-polymerase chain reaction; suberic bis-hydroxamic acid; suberoylanilide hydroxamic acid