title: [Effects of glycogen synthase kinase 3β overexpression in rat and glycogen synthase kinase 3β inhibitor SB-216763 on proliferation of hepatic oval cells].
abstract: OBJECTIVE
  To research the effects of glycogen synthase kinase (GSK3β) overexpression and GSK3β inhibitor SB-216763 on the proliferation of hepatic oval cells in rats and its regulatory mechanisms by Wnt signaling pathway.
  
  
  METHODS
  The hepatic oval cells WBF-344 were divided into the blank control group, GSK3β over-expression group, DMSO control group and GSK3β inhibitor groups, while the inhibitor groups set up three concentration gradients, that was 1, 5, 10 µmol/L. Using the GSK3β over-expression lentivirus, which had been identified correctly, and SB-216763 dealt with the cells WBF-344. The cells morphology of each group was observed under the phase contrast inverted microscope, and the expression of fluorescence in the lentivirus-transfected group was observed under the fluorescent microscope. The proliferation of each group cells was tested by CCK8 kits. The cells' apoptosis was detected by AnnexinV-FITC/PI kits. The expression of GSK3β, β-catenin and cyclin D1 were detected by Western blot.
  
  
  RESULTS
  The cells of GSK3β over-expression group were fewer and obvious aging. However, in each inhibitor added group, the cells' division and proliferation was vigorous, and the condition was good. Moreover, the cells' proliferation was getting stronger with the concentration of SB-216763 increasing. A large number of green fluorescence was expressed in the lentivirus-transfected cells. The cells' proliferation in GSK3β over-expression group restrained (t = 7.178, P < 0.01, as compared with control), while the cells' proliferation was vigorous in inhibitor groups (F = 45.030, P < 0.01, as compared with control). Flow Cytometry showed that the cells apoptosis was significant in GSK3β over-expression group. Western blot showed that the expression of GSK3β was increased, while the expression of β-catenin and cyclin D1 was decreased in the over-expression group. The expression of GSK3β had no significant difference among the control group and inhibitor groups. However, the expression of β-catenin and cyclin D1 was significantly increased with the concentration of SB-216763 increasing.
  
  
  CONCLUSIONS
  The overexpression of GSK3β can inhibit the Wnt signaling pathway, thus restrain the cells' proliferation and promotes apoptosis. SB-216763 can activate the Wnt pathway, thus promotes cells' proliferation.
authors.affiliation: Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, China.
authors.forename: Jun-qiao
authors.initials: JQ
authors.lastname: Zhong
authors.forename: Yuan-kang
authors.initials: YK
authors.lastname: Xie
authors.forename: Xiao-ke
authors.initials: XK
authors.lastname: Ji
authors.forename: Jun-hui
authors.initials: JH
authors.lastname: Fu
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Qi-yu
authors.initials: QY
authors.lastname: Zhang
authors.forename: Hong-qi
authors.initials: HQ
authors.lastname: Shi
authors.forename: Yun-feng
authors.initials: YF
authors.lastname: Shan
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D019938:Cyclin D1 / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D038341:Glycogen Synthase Kinases / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects*; D007211:Indoles / Q000494:pharmacology*; D008297:Male; D008301:Maleimides / Q000494:pharmacology*; D051381:Rats; D014162:Transfection; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism
chemical_list: C530106:Ccnd1 protein, rat; C495268:Ctnnb1 protein, rat; D007211:Indoles; D008301:Maleimides; C417521:SB 216763; D051176:beta Catenin; D019938:Cyclin D1; D038341:Glycogen Synthase Kinases; D000071679:Glycogen Synthase Kinase 3 beta; C000605508:Gsk3b protein, rat; D038362:Glycogen Synthase Kinase 3