title: Aberrant reduction of MiR-141 increased CD47/CUL3 in Hirschsprung's disease.
abstract: BACKGROUND
  MiR-141 has been confirmed to be associated with various human diseases. However, whether miR-141 is involved in the pathogenesis of Hirschsprung's disease (HSCR) remains unknown. Here, we design the experiment to reveal the relationship between miR-141 and HSCR.
  
  
  METHODS
  Quantitative real-time PCR and Western blot were used to detect the expression levels of miR-141 and its potential genes in 70 tissues of HSCR compared with 60 controls. Bisulfite sequencing PCR (BSP) assay was applied to explain the possible mechanism of the aberrant expression level of miR-141. We employed a dual-luciferase reporter assay to validate the regulation relation between miR-141 and CD47/CUL3. Cell migration, proliferation, apoptosis, and cell cycle progression were examined by transwell assay, MTT assay, and flow cytometry, respectively.
  
  
  RESULTS
  MiR-141 was down-regulated whereas CD47 and CUL3 expression was increased in colon tissues from patients with HSCR compared with control group, The increased level of CD47 and CUL3 induced by miR-141 reduced proliferation and migration of 293T and SH-SY5Y cells. Furthermore, this suppression was reversed by reducing of CD47 and CUL3. Hypermethylation of a CpG Island in the promoter region of miR-141 gene was confirmed in HSCR tissues.
  
  
  CONCLUSIONS
  Aberrant reduction of miR-141 may play an important role in the pathogenesis of HSCR with the inhibiting affection on cell migration and proliferation abilities. The present study demonstrates for the first time the role of miR-141 and its target genes in the occurrence of HSCR, and provides us a new direction for the study of the pathogenesis of Hirschsprung's disease.
authors: State Key Laboratory of Reproductive Medicine, Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing, China.
authors: Weibing
authors: W
authors: Tang
authors: Jingjing
authors: J
authors: Qin
authors: Junwei
authors: J
authors: Tang
authors: Hongwei
authors: H
authors: Zhang
authors: Zhigang
authors: Z
authors: Zhou
authors: Bo
authors: B
authors: Li
authors: Qiming
authors: Q
authors: Geng
authors: Wei
authors: W
authors: Wu
authors: Yankai
authors: Y
authors: Xia
authors: Xiaoqun
authors: X
authors: Xu
mesh_terms: D001483:Base Sequence; D051928:CD47 Antigen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D003106:Colon / Q000378:metabolism; D044842:Cullin Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D015536:Down-Regulation; D005260:Female; D057809:HEK293 Cells; D006627:Hirschsprung Disease / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007223:Infant; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000378:metabolism; D016415:Sequence Alignment; D015854:Up-Regulation
chemical_list: D051928:CD47 Antigen; C503645:CUL3 protein, human; D044842:Cullin Proteins; C533895:MIRN141 microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering