Protein kinase C delta inhibits the proliferation of vascular smooth muscle cells by suppressing G1 cyclin expression.
To elucidate the physiological role of protein kinase C (PKC) delta, a ubiquitously expressed isoform in vascular smooth muscle cells (VSMC), PKC delta was stably overexpressed in A7r5 cells, rat clonal VSMC. The [3H]thymidine incorporation in A7r5 overexpressed with PKC delta (DVs) was suppressed to 37.1 +/- 16.3% (mean +/- S.D.) of the level in control or A7r5 transfected with vector alone (EVs). The reduction of [3H]thymidine incorporation was strongly correlated with overexpressed PKC levels. Moreover, transient transfection of a dominant negative mutant of PKC delta restored the reduced proliferation in DVs. Flow cytometry analysis demonstrated that DVs were arrested in the G0/G1 phase of the cell cycle. Expression of cyclins D1 and E and retinoblastoma protein phosphorylation were reduced, while the protein levels of p27 were elevated in DVs as compared with EVs. There were no significant differences in the expression of c-fos, c-jun, c-myc, cyclin D2, D3, cyclin-dependent kinase 2, cyclin-dependent kinase 4, and p21 among the clones. We conclude that PKC delta inhibits the proliferation of VSMC by arresting cells in G1 via mainly inhibiting the expression of cyclin D1 and cyclin E.
Fukumoto
S
S
Second Department of Internal Medicine, Osaka City University Medical School, Osaka 545, Japan.
Nishizawa
Y
Y
Hosoi
M
M
Koyama
H
H
Yamakawa
K
K
Ohno
S
S
Morii
H
H
D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D056746:Cyclin G; D056766:Cyclin G1; D016213:Cyclins / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication; D003609:Dactinomycin / Q000494:pharmacology; D016193:G1 Phase; D007527:Isoenzymes / Q000494:pharmacology*; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D010766:Phosphorylation; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011493:Protein Kinase C / Q000494:pharmacology*; D051745:Protein Kinase C-delta; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D016160:Retinoblastoma Protein / Q000378:metabolism
C534704:Ccng1 protein, rat; D056746:Cyclin G; D056766:Cyclin G1; D016213:Cyclins; D007527:Isoenzymes; D010982:Platelet-Derived Growth Factor; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D016160:Retinoblastoma Protein; D003609:Dactinomycin; C497260:Prkcd protein, rat; D011493:Protein Kinase C; D051745:Protein Kinase C-delta