[Blockage of U251 cells in G0/G1 through MAPK signaling pathway by LRRC4].
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.
Wu
Ming-Hua
MH
Cancer Research Institute, Central South University, Changsha, China.
Huang
Chen
C
Li
Xiao-Ling
XL
Zhou
Ming
M
Zhou
Yan-Hong
YH
Zhang
Zu-Ping
ZP
Li
Gui-Yuan
GY
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
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