title: SOX4 interacts with EZH2 and HDAC3 to suppress microRNA-31 in invasive esophageal cancer cells.
abstract: BACKGROUND
  Tumor metastasis is responsible for 90% of cancer-related deaths. Recently, a strong link between microRNA dysregulation and human cancers has been established. However, the molecular mechanisms through which microRNAs regulate metastasis and cancer progression remain unclear.
  
  
  METHODS
  We analyzed the reciprocal expression regulation of miR-31 and SOX4 in esophageal squamous and adenocarcinoma cell lines by qRT-PCR and Western blotting using overexpression and shRNA knock-down approaches. Furthermore, methylation studies were used to assess epigenetic regulation of expression. Functionally, we determined the cellular consequences using migration and invasion assays, as well as proliferation assays. Immunoprecipitation and ChIP were used to identify complex formation of SOX4 and co-repressor components.
  
  
  RESULTS
  Here, we report that SOX4 promotes esophageal tumor cell proliferation and invasion by silencing miR-31 via activation and stabilization of a co-repressor complex with EZH2 and HDAC3. We demonstrate that miR-31 is significantly decreased in invasive esophageal cancer cells, while upregulation of miR-31 inhibits growth, migration and invasion of esophageal adenocarcinoma (EAC) and squamous cell carcinoma (ESCC) cell lines. miR-31, in turn, targets SOX4 for degradation by directly binding to its 3'-UTR. Additionally, miR-31 regulates EZH2 and HDAC3 indirectly. SOX4, EZH2 and HDAC3 levels inversely correlate with miR-31 expression in ESCC cell lines. Ectopic expression of miR-31 in ESCC and EAC cell lines leads to down regulation of SOX4, EZH2 and HDAC3. Conversely, pharmacologic and genetic inhibition of SOX4 and EZH2 restore miR-31 expression. We show that SOX4, EZH2 and HDAC3 form a co-repressor complex that binds to the miR-31 promoter, repressing miR-31 through an epigenetic mark by H3K27me3 and by histone acetylation. Clinically, when compared to normal adjacent tissues, esophageal tumor samples show upregulation of SOX4, EZH2, and HDAC3, and EZH2 expression is significantly increased in metastatic ESCC tissues.
  
  
  CONCLUSIONS
  Thus, we identified a novel molecular mechanism by which the SOX4, EZH2 and miR-31 circuit promotes tumor progression and potential therapeutic targets for invasive esophageal carcinomas.
authors: Department of Surgery, 2213 Garland Ave. 10445 MRB IV, Nashville, TN, 37232-6840, USA. rainelli.koumangoye@vanderbilt.edu.
authors: Rainelli B
authors: RB
authors: Koumangoye
authors: Division of Dermatology, Department of Medicine, 21st Ave South, A-2310 Medical Center North, Nashville, TN, 37232-6840, USA. thomas.andl@vanderbilt.edu.
authors: Thomas
authors: T
authors: Andl
authors: Department of Surgery, 2213 Garland Ave. 10445 MRB IV, Nashville, TN, 37232-6840, USA. kenneth.j.taubenslag@Vanderbilt.Edu.
authors: Kenneth J
authors: KJ
authors: Taubenslag
authors: Department of Surgery, 2213 Garland Ave. 10445 MRB IV, Nashville, TN, 37232-6840, USA. steven.t.zilberman@Vanderbilt.Edu.
authors: Steven T
authors: ST
authors: Zilberman
authors: Department of Surgery, 2213 Garland Ave. 10445 MRB IV, Nashville, TN, 37232-6840, USA. claudia.andl@vanderbilt.edu.
authors: Chase J
authors: CJ
authors: Taylor
authors: Department of Cancer Biology, 2213 Garland Ave. 10445 MRB IV, Nashville, TN, 37232-6840, USA. holli.a.loomans@vanderbilt.edu.
authors: Holli A
authors: HA
authors: Loomans
authors: Department of Surgery, 2213 Garland Ave. 10445 MRB IV, Nashville, TN, 37232-6840, USA. claudia.andl@vanderbilt.edu.|Department of Cancer Biology, 2213 Garland Ave. 10445 MRB IV, Nashville, TN, 37232-6840, USA. claudia.andl@vanderbilt.edu.|Vanderbilt Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN, 37232-6840, USA. claudia.andl@vanderbilt.edu.|Vanderbilt Digestive Disease Center, Vanderbilt University Medical Center, Nashville, TN, 37232-6840, USA. claudia.andl@vanderbilt.edu.
authors: Claudia D
authors: CD
authors: Andl
mesh_terms: D020413:3' Untranslated Regions; D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D015536:Down-Regulation; D000071221:Enhancer of Zeste Homolog 2 Protein; D044127:Epigenesis, Genetic; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D009361:Neoplasm Invasiveness; D063151:Polycomb Repressive Complex 2 / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D034622:RNA Interference; D012097:Repressor Proteins / Q000378:metabolism; D055751:SOXC Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000099186:Histone Deacetylase 3
chemical_list: D020413:3' Untranslated Regions; D000071221:Enhancer of Zeste Homolog 2 Protein; D006655:Histone Deacetylases; D035683:MicroRNAs; D063151:Polycomb Repressive Complex 2; D012097:Repressor Proteins; D055751:SOXC Transcription Factors; D000099186:Histone Deacetylase 3; C534612:MIRN31 microRNA, human; C083431:SOX4 protein, human; C498409:EZH2 protein, human
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