Acute glucose-induced downregulation of PKC-betaII accelerates cultured VSMC proliferation.
Accelerated vascular smooth muscle cell (VSMC) proliferation contributes to the formation of atherosclerotic lesions. To investigate protein kinase C (PKC)-betaII functions with regard to glucose-induced VSMC proliferation, human VSMC from aorta (AoSMC), a clonal VSMC line of rat aorta (A10), and A10 cells overexpressing PKC-betaI (betaI-A10) and PKC-betaII (betaII-A10) were studied with the use of three techniques to evaluate glucose effects on aspects affecting proliferation. High glucose (25 mM) increased DNA synthesis and accelerated cell proliferation compared with normal glucose (5.5 mM) in AoSMC and A10 cells, but not in betaI-A10 and betaII-A10 cells. The PKC-betaII specific inhibitor CGP-53353 inhibited glucose-induced cell proliferation and DNA synthesis in AoSMC and A10 cells. In flow cytometry analysis, high glucose increased the percentage of A10 cells at 12 h after cell cycle initiation but did not increase the percentage of betaI-A10 or betaII-A10 cells entering S phase. PKC-betaII protein levels decreased before the peak of DNA synthesis, and high glucose further decreased PKC-betaII mRNA and protein levels in AoSMC and A10 cells. These results suggest that high glucose downregulates endogenous PKC-betaII, which then alters the normal inhibitory role of PKC-betaII in cell cycle progression, resulting in the stimulation of VSMC proliferation through acceleration of the cell cycle.
Yamamoto
M
M
Department of Biochemistry and Molecular Biology, University of South Florida, College of Medicine, Tampa, Florida 33612, USA.
Acevedo-Duncan
M
M
Chalfant
C E
CE
Patel
N A
NA
Watson
J E
JE
Cooper
D R
DR
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D015536:Down-Regulation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D010797:Phthalimides / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D064546:Protein Kinase C beta; D051381:Rats; D016196:S Phase / Q000187:drug effects; D013936:Thymidine / Q000037:antagonists & inhibitors / Q000378:metabolism; D013997:Time Factors
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D010797:Phthalimides; D004247:DNA; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D005947:Glucose; D013936:Thymidine; C098874:4,5-dianilinophthalimide