title: Expression and role of oncogenic miRNA-224 in esophageal squamous cell carcinoma.
abstract: BACKGROUND
  Aberrant expression of miR-224 is associated with tumor development and progression. This study investigated the role of miR-224 in esophageal squamous cell carcinoma (ESCC) ex vivo and in vitro.
  
  
  METHODS
  A total of 103 esophageal intraepithelial neoplasia, ESCC tissue specimens, and their matched distant normal tissues were collected to test miR-224 expression using qRT-PCR analysis. Western blot was used to quantify the level of PH domain leucine-rich repeat protein phosphatase 1 (PHLPP1) and PHLPP2 in ESCC tissues. Cell viability, apoptosis, invasion, and colony formation assays were used to assess the altered phenotypes of esophageal cancer cell lines after miR-224 expression or inhibition. A luciferase reporter assay was used to confirm miR-224 binding to PHLPP1 and PHLPP2 mRNA.
  
  
  RESULTS
  miR-224 was significantly overexpressed in esophageal intraepithelial neoplasia and ESCC tissues, while the expression of PHLPP1 and PHLPP2 proteins, the target genes of miR-224, was downregulated in ESCC tissues. miR-224 expression was associated with advanced clinical TNM stage, pathologic grade, and the level of PHLPP1 and PHLPP2 proteins in ESCC tissues. Ectopic overexpression of miR-224 promoted proliferation, migration, and invasion, but suppressed apoptosis of ESCC cells. miR-224 was able to bind to the 3' untranslated region (3'-UTR) of PHLPP1 and PHLPP2 mRNA to suppress their expression.
  
  
  CONCLUSIONS
  The current study demonstrated that miR-224 acts as an oncogenic miRNA in ESCC, possibly by targeting PHLPP1 and PHLPP2.
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. xiaoyanhe1988@126.com.|Department of Gastroenterology, Dongyang People's Hospital, 60 Wuningxi Road, Jinhua, China. xiaoyanhe1988@126.com.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: He
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. 1365360153@qq.com.|Department of Gastroenterology, The First People's Hospital of Lianyungang, 182 Tongguanbei Road, Lianyungang, China. 1365360153@qq.com.
authors.forename: Zhimei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. limingibm@sina.com.|Department of Gastroenterology, Friendship Hospital of Yangzhou, 440 Siwangting Road, Yangzhou, China. limingibm@sina.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. lishuo1989@163.com.
authors.forename: Shuo
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. renlihuaxy@126.com.|Department of Gastroenterology, Zhangjiagang First People's Hospital, 68 Jiyangxi Road, Suzhou, China. renlihuaxy@126.com.
authors.forename: Lihua
authors.initials: L
authors.lastname: Ren
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. zhuhong1059@126.com.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. lancent@gmail.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Xiao
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, China. ruihuashi@126.com.|Department of Gastroenterology, Zhongda Hospital, Southeast University, 87 Dingjiaqiao Road, Nanjing, China. ruihuashi@126.com.
authors.forename: Ruihua
authors.initials: R
authors.lastname: Shi
mesh_terms: D020413:3' Untranslated Regions; D000328:Adult; D000368:Aged; D017209:Apoptosis; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement; D002470:Cell Survival; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000077277:Esophageal Squamous Cell Carcinoma; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; C546140:MIRN224 microRNA, human; D035683:MicroRNAs; D009687:Nuclear Proteins; C400601:PHLPP1 protein, human; C541572:PHLPP2 protein, human; D010749:Phosphoprotein Phosphatases