title: Hepatocyte nuclear factor 6 inhibits the growth and metastasis of cholangiocarcinoma cells by regulating miR-122.
abstract: OBJECTIVE
  Hepatocyte nuclear factor 6 (HNF6) is a liver-enriched transcription factor and highly expressed in mature bile duct epithelial cells. This study sought to investigate the role of HNF6, particularly the molecular mechanisms for how HNF6 is involved in the growth and metastasis of cholangiocarcinoma (CCA) cells.
  
  
  METHODS
  The expression of HNF6, miR-122 and key molecules was examined by Western blot analysis and real-time RT-PCR. Stable transfectants, HCCC-HNF(low) and RBE-HNF(high), were generated from human CCA HCCC-9810 and RBE cells, respectively. The regulatory effect of HNF6 on miR-122 was evaluated by luciferase reporter assay. Cell proliferation, cycle distribution, migration and invasion were analyzed. The xenograft model was used to assess the effects of HNF6 overexpression on tumorigenesis, growth, metastasis and therapeutic potentials.
  
  
  RESULTS
  Human CCA tissues and cells expressed lower levels of HNF6, which positively correlated with miR-122. HNF6 regulated the expression of miR-122 by stimulating its promoter. HNF6 overexpression inhibited cell proliferation by inducing cell cycle arrest at G1 phase through regulating miR-122, cyclin G1 and insulin-like growth factor-1 receptor. HNF6 inhibited the migration and invasion of CCA cells by regulating matrix metalloproteinase-2 and metalloproteinase-9, reversion-inducing-cysteine-rich protein with kazal motifs, E-cadherin and N-cadherin. Co-transfection of anti-miR-122 abrogated the effects of HNF6. HNF6 overexpression inhibited the ability of cells to form tumors and to metastasize to the lungs of mice, and the growth of established tumors.
  
  
  CONCLUSIONS
  The results indicate that HNF6 may serve as a tumor suppressor by regulating miR-122, and its overexpression may represent a mechanism-based therapy for CCA.
authors.affiliation: Department of General Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China.
authors.forename: Huaqiang
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of General Surgery, Liaocheng People's Hospital, Liaocheng, 252000, China.
authors.forename: Yuetang
authors.initials: Y
authors.lastname: Mi
authors.affiliation: Key Laboratory of Hepatosplenic Surgery, Department of General Surgery, First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China.
authors.forename: Xian
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Department of General Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of General Surgery, Liaocheng People's Hospital, Liaocheng, 252000, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.affiliation: Key Laboratory of Hepatosplenic Surgery, Department of General Surgery, First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wei
authors.affiliation: Key Laboratory of Hepatosplenic Surgery, Department of General Surgery, First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China.
authors.forename: Hongchi
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of General Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Lu
authors.affiliation: Key Laboratory of Hepatosplenic Surgery, Department of General Surgery, First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China. kevsun88@hotmail.com.|Department of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, 1005, New Zealand. kevsun88@hotmail.com.
authors.forename: Xueying
authors.initials: X
authors.lastname: Sun
mesh_terms: D000818:Animals; D001650:Bile Duct Neoplasms / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control* / Q000556:secondary; D001653:Bile Ducts, Intrahepatic / Q000378:metabolism* / Q000473:pathology; D015153:Blotting, Western; D002453:Cell Cycle; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D018281:Cholangiocarcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D015972:Gene Expression Regulation, Neoplastic; D051559:Hepatocyte Nuclear Factor 6 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
chemical_list: D051559:Hepatocyte Nuclear Factor 6; C531907:MIRN122 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger
keywords: Cholangiocarcinoma; Hepatocyte nuclear factor 6; Metastasis; MicroRNA-122; Proliferation
references.citation: J Clin Invest. 2012 Aug;122(8):2884-97
references.pmid: 22820290
references.citation: PLoS One. 2010 Oct 13;5(10):e13344
references.pmid: 20967225
references.citation: Cell. 2001 Dec 14;107(6):789-800
references.pmid: 11747814
references.citation: J Clin Invest. 2012 Aug;122(8):2871-83
references.pmid: 22820288
references.citation: Semin Cancer Biol. 2010 Jun;20(3):161-8
references.pmid: 20470890
references.citation: Cell. 2007 Jun 29;129(7):1401-14
references.pmid: 17604727
references.citation: Oncogene. 2009 Oct 8;28(40):3526-36
references.pmid: 19617899
references.citation: Cancer Biol Ther. 2010 Apr 1;9(7):554-61
references.pmid: 20150764
references.citation: N Engl J Med. 2010 Apr 8;362(14):1273-81
references.pmid: 20375404
references.citation: Genes Dev. 2006 Aug 15;20(16):2293-305
references.pmid: 16912278
references.citation: Hepatology. 2009 May;49(5):1571-82
references.pmid: 19296470
references.citation: J Hepatol. 2014 Jun;60(6):1268-89
references.pmid: 24681130
references.citation: Cell Signal. 2013 Apr;25(4):931-8
references.pmid: 23333463
references.citation: Mol Cancer. 2015 Aug 25;14:163
references.pmid: 26302777
references.citation: Liver Int. 2014 Feb;34(2):281-95
references.pmid: 23834235
references.citation: Cell Signal. 2012 Jan;24(1):9-16
references.pmid: 21893194
references.citation: Mol Carcinog. 2013 Apr;52(4):297-303
references.pmid: 22213236
references.citation: FEBS J. 2014 Jan;281(1):115-28
references.pmid: 24165223
references.citation: J Biol Chem. 2015 Jan 9;290(2):1170-85
references.pmid: 25422324
references.citation: Hepatology. 2008 Mar;47(3):897-907
references.pmid: 18176954
references.citation: Hepatology. 2010 Nov;52(5):1702-12
references.pmid: 21038412
references.citation: FASEB J. 2007 May;21(7):1445-62
references.pmid: 17283222
references.citation: PLoS One. 2012;7(10):e47053
references.pmid: 23056576
references.citation: Hepatology. 2010 Oct;52(4):1431-42
references.pmid: 20842632
references.citation: Cancer Res. 2007 Jul 1;67(13):6092-9
references.pmid: 17616664
references.citation: Gastroenterology. 2012 Jan;142(1):119-29
references.pmid: 21920465
references.citation: J Hepatol. 2015 Feb;62(2):448-57
references.pmid: 25308172
references.citation: Nat Rev Gastroenterol Hepatol. 2011 Aug 02;8(9):512-22
references.pmid: 21808282
references.citation: Lab Invest. 2014 May;94(5):517-27
references.pmid: 24638272
references.citation: Oncotarget. 2015 Oct 6;6(30):28867-81
references.pmid: 26311740
references.citation: Science. 2004 Mar 5;303(5663):1483-7
references.pmid: 15001769
references.citation: PLoS One. 2014 Jul 03;9(7):e101330
references.pmid: 24992599
references.citation: J Biol Chem. 2013 Oct 25;288(43):31206-16
references.pmid: 24022481
references.citation: Mol Cell Biochem. 2015 Feb;400(1-2):1-7
references.pmid: 25472877
references.citation: Drug Metab Pharmacokinet. 2013;28(3):250-9
references.pmid: 23291635