title: [Role of miR-21 in rat hepatic oval cell proliferation and activation].
abstract: OBJECTIVE
  To investigate the possible mechanisms of miR-21-mediated regulation of proliferation and activation of hepatic oval cells.
  
  
  METHODS
  The 2-acetamidofluorene/partial hepatectomy (2-AAF/PH) method was applied to generate hepatic oval cell activation model in male Sprague-Dawley rats; after the 7 days of 2-AAF/PH or PH alone (control), the rats were sacrificed at 0 h, 6 h, 12 h, 24 h, 72 h and 168 h. Expression of miR-21 was detected by real-time PCR and differences between groups were evaluated using the two-sample t-test. Differential transcription of miR-21 target genes was assessed bioinformatically, and with western blotting to detect changes in protein expression of the target gene.
  
  
  RESULTS
  The rat hepatic oval cell activation model was successfully established.The 2-AAF/PH rats showed miR-21 expression beginning to increase at 12 h, peaking at 24 h, and decreasing thereafter until an increase at 168 h.For the control group, the miR-21 expression began to increase at 6 h, until 24 h when expression began steadily declining to reach the original level.Compared to the control group, the experimental group showed expression of miR-21 that was significantly less at 6 h (P=0.039, t =3.029) and significantly more at 24 h and 168 h (P=0.026, t =-3.433 and P=0.007, t =-5.105). Among the predicted target genes of miR-21 were WW domain containing E3 ubiquitin protein ligase 1 (WWD), Smad family member 7 (Smad7), and polybromo-1 (Pbrm1).Smad7 protein expression began to decrease at 6 h in the control group, until reaching its minimum at 24 h when it increased; in the experimental group, SMAD7 expression increased at 6 h, then began to decrease with the minimum detected at 168 hour.In the control group, the Smad7 mRNA expression decreased slightly at 6 h, then began to increase, reaching its peak at 24 h when the expression fell to the original level. In the experimental group, the Smad7 mRNA expression began to increase at 6 h and reached its peak at 24 h when it decreased; the expression was little more than its original level at 168 h.Smad7 protein expression was negatively correlated with miR-21, and Smad7 mRNA expression was positively correlated with miR-21 but negatively correlated with Smad7 protein expression.
  
  
  CONCLUSIONS
  miR-21 may play a vital role in the activation and proliferation of hepatic oval cells.As a target gene of miR-21, Smad7 might be involved in the process.
authors.affiliation: Department of General Surgery 2, the First People's Hospital of Neijiang, Neijiang 641000, China.
authors.forename: Zhen-Kun
authors.initials: ZK
authors.lastname: Chen
authors.forename: Yun-Feng
authors.initials: YF
authors.lastname: Shan
authors.forename: Bin
authors.initials: B
authors.lastname: He
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.forename: Bo
authors.initials: B
authors.lastname: Wu
authors.forename: Xiao-Ke
authors.initials: XK
authors.lastname: Ji
authors.forename: Si-Lu
authors.initials: SL
authors.lastname: Wang
authors.forename: Qi-Yu
authors.initials: QY
authors.lastname: Zhang
mesh_terms: D015073:2-Acetylaminofluorene; D000818:Animals; D049109:Cell Proliferation; D006498:Hepatectomy; D022781:Hepatocytes / Q000166:cytology*; D008099:Liver / Q000166:cytology*; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D035683:MicroRNAs; D015073:2-Acetylaminofluorene