Down-regulation of protein kinase C alpha and gamma and enhanced TPA-induced neurite formation in DAN-transfected neuroblastoma cells.
DAN gene was first isolated by differential screening between rat 3Y1 and v-src-transformed 3Y1 cells and showed a tumor-suppressive activity toward v-src-transformed 3Y1 cells. When DAN-transfected neuroblastoma cells were treated with a tumor promoter phorbol ester, TPA, neurite-like processes appeared within 2 h whereas no apparent change was observed in the parent and vector-transfected cells up to 8 h. This suggests some difference in TPA-receptor, protein kinase C (PKC), between DAN-transfectants and the control cells. DAN-transfected SH-SY5Y cells showed complete loss in PKCalpha and a large decrease in PKCgamma. Similar down-regulation in PKCalpha and PKCgamma was also observed in DAN-transfected Ha-ras-transformed NIH 3T3 cells. The decreased level of PKCalpha was partially recovered after treatment with a calpain inhibitor, ZLLH. A 150-kDa proteolytic product of a calpain-specific substrate, non-erythroid alpha-spectrin, was detectable in DAN-transfected SH-SY5Y cells but not in the parent or vector-transfected control cells. This suggests that DAN-transfected cells contain activated calpain which may cause down-regulation of PKC and hence induce the altered TPA response.
Hiwasa
T
T
Department of Biochemistry, School of Medicine, Chiba University, Japan. hiwasa@med.m.chiba-u.ac.jp
Nakamura
Y
Y
Ozaki
T
T
Kondo
K
K
Saido
T
T
Nakagawara
A
A
Sakiyama
S
S
D016475:3T3 Cells; D000818:Animals; D015153:Blotting, Western; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins; D016207:Cytokines; D003907:Dexamethasone; D015536:Down-Regulation; D005347:Fibroblasts; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D008970:Molecular Weight; D009419:Nerve Tissue Proteins; D016501:Neurites / Q000187:drug effects* / Q000378:metabolism; D009447:Neuroblastoma; D009474:Neurons / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014162:Transfection; D014407:Tumor Cells, Cultured; D013049:Spectrin
D002154:Calpain; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D016207:Cytokines; D003907:Dexamethasone; D007527:Isoenzymes; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D011506:Proteins; D013755:Tetradecanoylphorbol Acetate; D013049:Spectrin; C490088:Grem2 protein, mouse; C490087:NBL1 protein, human; C490089:Nbl1 protein, mouse; C085763:Nbl1 protein, rat; C087857:protein kinase C gamma; C496628:PRKCA protein, human; C496630:Prkca protein, mouse