title: miRNA-200c inhibits invasion and metastasis of human non-small cell lung cancer by directly targeting ubiquitin specific peptidase 25.
abstract: BACKGROUND
  Growing evidence indicates that miR-200c is involved in carcinogenesis and tumor progression in non-small-cell lung cancer (NSCLC). However, its precise biological role remains largely elusive.
  
  
  METHODS
  The functions of miR-200c and USP25 in migration/invasion and lung metastasis formation were determined by transwell and tail vein injection assays, respectively. The potential regulatory targets of miR-200c were determined by prediction tools, correlation with target protein expression, and luciferase reporter assay. The mRNA expression levels of miR-200c and USP25 were examined in NSCLC cell lines and patient specimens using quantitative reverse transcription-PCR. The protein expression levels of USP25 were examined in NSCLC cell lines and patient specimens using western blot and immunohistochemical staining.
  
  
  RESULTS
  We demonstrated that over-expression of miR-200c inhibited NSCLC cells migration, invasion, epithelial-mesenchymal transition (EMT) in vitro and lung metastasis formation in vivo. Further studies revealed that USP25 was a downstream target of miR-200c in NSCLC cells as miR-200c bound directly to the 3'-untranslated region of USP25, thus reducing both the messenger RNA and protein levels of USP25. Silencing of the USP25 gene recapitulated the effects of miR-200c over-expression. Clinical analysis indicated that miR-200c was negatively correlated with clinical stage, lymph node metastasis in NSCLC patients. Moreover, USP25 protein and mRNA level expressions were higher in NSCLC patients, compared to healthy control, and correlated with clinical stage and lymphatic node metastasis.
  
  
  CONCLUSIONS
  These findings indicate that miR-200c exerts tumor-suppressive effects for NSCLC through the suppression of USP25 expression and suggests a new therapeutic application of miR-200c in the treatment of NSCLC.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Qiang
authors.initials: Q
authors.lastname: Tan
authors.forename: Mingxia
authors.initials: M
authors.lastname: Yan
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
authors.forename: Hechun
authors.initials: H
authors.lastname: Lin
authors.forename: Fangyu
authors.initials: F
authors.lastname: Zhao
authors.forename: Guoliang
authors.initials: G
authors.lastname: Bao
authors.forename: Hanwei
authors.initials: H
authors.lastname: Kong
authors.forename: Chao
authors.initials: C
authors.lastname: Ge
authors.forename: Fanglin
authors.initials: F
authors.lastname: Zhang
authors.forename: Tao
authors.initials: T
authors.lastname: Yu
authors.forename: Jinjun
authors.initials: J
authors.lastname: Li
authors.forename: Xianghuo
authors.initials: X
authors.lastname: He
authors.affiliation: State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 200032 Shanghai, China. myao@shsci.org.
authors.forename: Ming
authors.initials: M
authors.lastname: Yao
mesh_terms: D000368:Aged; D000818:Animals; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D008207:Lymphatic Metastasis; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D009361:Neoplasm Invasiveness / Q000235:genetics / Q000473:pathology; D043222:Ubiquitin Thiolesterase / Q000235:genetics*
chemical_list: C533896:MIRN200 microRNA, human; D035683:MicroRNAs; C404488:USP25 protein, human; D043222:Ubiquitin Thiolesterase