title: [Blockage of U251 cells in G0/G1 through MAPK signaling pathway by LRRC4].
abstract: OBJECTIVE
  To explore the effect of LRRC4, a glioma suppressive gene, on blocking U251 cells in G0/G1 by MAPK signaling pathway.
  
  
  METHODS
  LRRC4 was transfected into U251 cells, and at 24 hour of post-transfection, cells were split at a 1:3 dilution, challenged with 500 microg /mL G418 and formed a stable transfected clone pool. RT-PCR, Northern blot and Western blot were used to identify the stable transfectants. ERK, JNK and P38 expression changes were analyzed by Western blot. FACS analysis, Luciferase reporter gene assay and Western blot were used to detect the cell cycle and cyclin D1.
  
  
  RESULTS
  LRRC4 down-regulated the expression of phosphorylated ERK2 and up-regulated the expression of total protein JNK2 (a key molecule of MAPK signaling pathway) and phosphorylated c-Jun. LRRC4 decreased the expression of mutation P53, cyclin D1 activation and its expression. U251 cells were blocked in G0/G1 by LRRC4.
  
  
  CONCLUSIONS
  LRRC4 can decrease JNK2, up-regulate the phosphoralated c-Jun, down-regulate mutant P53 and cyclin D1, and therefore block U251 cells in G0/G1.
authors.affiliation: Cancer Research Institute, Central South University, Changsha, China.
authors.forename: Ming-Hua
authors.initials: MH
authors.lastname: Wu
authors.forename: Chen
authors.initials: C
authors.lastname: Huang
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Zhou
authors.forename: Yan-Hong
authors.initials: YH
authors.lastname: Zhou
authors.forename: Zu-Ping
authors.initials: ZP
authors.lastname: Zhang
authors.forename: Gui-Yuan
authors.initials: GY
authors.lastname: Li
mesh_terms: D015152:Blotting, Northern; D015153:Blotting, Western; D045744:Cell Line, Tumor; D019938:Cyclin D1 / Q000378:metabolism; D005434:Flow Cytometry; D016193:G1 Phase / Q000235:genetics / Q000502:physiology*; D005910:Glioma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000235:genetics / Q000502:physiology*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016192:Resting Phase, Cell Cycle / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: C503331:LRRC4 protein, human; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D019938:Cyclin D1; D008156:Luciferases; D020929:Mitogen-Activated Protein Kinase Kinases