title: MiRNA-205 modulates cellular invasion and migration via regulating zinc finger E-box binding homeobox 2 expression in esophageal squamous cell carcinoma cells.
abstract: BACKGROUND
  Esophageal squamous cell carcinoma (ESCC) is often diagnosed at later stages until they are incurable. MicroRNA (miR) is a small, non-coding RNA that negatively regulates gene expression mainly via translational repression. Accumulating evidence indicates that deregulation of miR is associated with human malignancies including ESCC. The aim of this study was to identify miR that could be specifically expressed and exert distinct biological actions in ESCC.
  
  
  METHODS
  Total RNA was extracted from ESCC cell lines, OE21 and TE10, and a non-malignant human esophageal squamous cell line, Het-1A, and subjected to microarray analysis. Expression levels of miR that showed significant differences between the 2 ESCC and Het-1A cells based on the comprehensive analysis were analyzed by the quantitative reverse transcriptase (RT)-PCR method. Then, functional analyses, including cellular proliferation, apoptosis and Matrigel invasion and the wound healing assay, for the specific miR were conducted. Using ESCC tumor samples and paired surrounding non-cancerous tissue obtained endoscopically, the association with histopathological differentiation was examined with quantitative RT-PCR.
  
  
  RESULTS
  Based on the miR microarray analysis, there were 14 miRs that showed significant differences (more than 2-fold) in expression between the 2 ESCC cells and non-malignant Het-1A. Among the significantly altered miRs, miR-205 expression levels were exclusively higher in 5 ESCC cell lines examined than any other types of malignant cell lines and Het-1A. Thus, miR-205 could be a specific miR in ESCC. Modulation of miR-205 expression by transfection with its precursor or anti-miR-205 inhibitor did not affect ESCC cell proliferation and apoptosis, but miR-205 was found to be involved in cell invasion and migration. Western blot revealed that knockdown of miR-205 expression in ESCC cells substantially enhanced expression of zinc finger E-box binding homeobox 2, accompanied by reduction of E-cadherin, a regulator of epithelial mesenchymal transition. The miR-205 expression levels were not associated with histological differentiation of human ESCC.
  
  
  CONCLUSIONS
  These results imply that miR-205 is an ESCC-specific miR that exerts tumor-suppressive activities with EMT inhibition by targeting ZEB2.
authors.affiliation: Department of Gastroenterology and Hepatology, Nagasaki University Hospital, Sakamoto, Nagasaki, Japan.
authors.forename: Kayoko
authors.initials: K
authors.lastname: Matsushima
authors.forename: Hajime
authors.initials: H
authors.lastname: Isomoto
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Yamaguchi
authors.forename: Naoki
authors.initials: N
authors.lastname: Inoue
authors.forename: Haruhisa
authors.initials: H
authors.lastname: Machida
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Nakayama
authors.forename: Tomayoshi
authors.initials: T
authors.lastname: Hayashi
authors.forename: Masaki
authors.initials: M
authors.lastname: Kunizaki
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Hidaka
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nagayasu
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nakashima
authors.forename: Kenta
authors.initials: K
authors.lastname: Ujifuku
authors.forename: Norisato
authors.initials: N
authors.lastname: Mitsutake
authors.forename: Akira
authors.initials: A
authors.lastname: Ohtsuru
authors.forename: Shunichi
authors.initials: S
authors.lastname: Yamashita
authors.forename: Manav
authors.initials: M
authors.lastname: Korpal
authors.forename: Yibin
authors.initials: Y
authors.lastname: Kang
authors.forename: Philip A
authors.initials: PA
authors.lastname: Gregory
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Goodall
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kohno
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Nakao
mesh_terms: D017209:Apoptosis / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000473:pathology; D002454:Cell Differentiation / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D049109:Cell Proliferation; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000473:pathology*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D010641:Phenotype; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics; D000074165:Zinc Finger E-box Binding Homeobox 2
chemical_list: D018398:Homeodomain Proteins; D035683:MicroRNAs; D012097:Repressor Proteins; D000074165:Zinc Finger E-box Binding Homeobox 2; C547183:MIRN10 microRNA, human; C534425:MIR205, human; C492363:ZEB2 protein, human