title: Decreased MiR-200a/141 suppress cell migration and proliferation by targeting PTEN in Hirschsprung's disease.
abstract: OBJECTIVE
  Hirschsprung's disease (HSCR) is a genetic disorder of neural crest development. In this study, we investigated whether and how miR-200a and miR-141, belonging to miR-200 family, were involved in the pathogenesis of HSCR.
  
  
  METHODS
  Quantitative real time PCR and Western blot were used to detect the levels of miRNA, mRNAs, and proteins in colon tissues from 88 HSCR patients and 75 controls. The direct regulation of specific mRNA by miRNAs was validated by dual-luciferase reporter assay and RNA interference in cell lines. Transwell assays, CCK8 assay, and flow cytometry were inplemented to measure viability and activities of human 293T and SH-SY5Y cells, respectively.
  
  
  RESULTS
  Aberrant suppression of miR-200a was observed in colon tissues of HSCR patients. A decreased level of miR-200a and miR-141 correlated with increased levels of PTEN mRNA and protein. The Dual-Luciferase reporter gene assay demonstrated that miR-200a and miR-141 binded directly to 3'UTR of PTEN and resulting in the inhibition of PTEN. The reductions in miR-200a and miR-141 inhibited migration and proliferation of 293T and SH-SY5Y cells through up-regulating the expression of PTEN. Moreover, knocking-down of PTEN rescued the extent of suppressed cell migration and proliferation induced by miR-200a and miR-141.
  
  
  CONCLUSIONS
  The miR-200 family may play a crucial role in the pathogenesis of HSCR by co-regulating PTEN.
authors.affiliation: State Key Laboratory of Reproductive Medicine, Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing, China.
authors.forename: Hongxing
authors.initials: H
authors.lastname: Li
authors.forename: Junwei
authors.initials: J
authors.lastname: Tang
authors.forename: Hao
authors.initials: H
authors.lastname: Lei
authors.forename: Peng
authors.initials: P
authors.lastname: Cai
authors.forename: Hairong
authors.initials: H
authors.lastname: Zhu
authors.forename: Bo
authors.initials: B
authors.lastname: Li
authors.forename: Xiaoqun
authors.initials: X
authors.lastname: Xu
authors.forename: Yankai
authors.initials: Y
authors.lastname: Xia
authors.forename: Weibing
authors.initials: W
authors.lastname: Tang
mesh_terms: D001483:Base Sequence; D002465:Cell Movement; D049109:Cell Proliferation; D017931:DNA Primers; D005260:Female; D057809:HEK293 Cells; D006627:Hirschsprung Disease / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007223:Infant; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D051059:PTEN Phosphohydrolase / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D015854:Up-Regulation
chemical_list: D017931:DNA Primers; C533895:MIRN141 microRNA, human; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human