Heparin selectively inhibits a protein kinase C-dependent mechanism of cell cycle progression in calf aortic smooth muscle cells.
The proliferation of arterial smooth muscle cells (SMCs) plays a critical role in the pathogenesis of arteriosclerosis. Previous studies have indicated that the glycosaminoglycan heparin specifically inhibited the growth of vascular SMCs in vivo and in culture, although the precise mechanism(s) of action have not been elucidated. In this study, we have examined the ability of specific mitogens (PDGF, EGF, heparin-binding growth factors, phorbol esters, and insulin) to stimulate SMC proliferation. Our results indicate that SMCs derived from different species and vascular sources respond differently to these growth factors. We next examined the ability of heparin to inhibit the proliferative responses to these mitogens. In calf aortic SMCs, heparin inhibits a protein kinase C-dependent pathway for mitogenesis. Detailed cell cycle analysis revealed several new features of the effects of heparin on SMCs. For example, heparin has two effects on the Go----S transition: it delays entry into S phase and also reduces the number of cells entering the cycle from Go. Using two separate experimental approaches, we found that heparin must be present during the last 4 h before S phase, suggesting a mid-to-late G1 heparin block. In addition, our data indicate that heparin-treated SMCs, while initially blocked in mid-to-late G1, slowly move back into a quiescent growth state in the continued presence of heparin. These results suggest that heparin may have multiple targets for its antiproliferative effect.
Castellot
J J
JJ
Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115.
Pukac
L A
LA
Caleb
B L
BL
Wright
T C
TC
Karnovsky
M J
MJ
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000187:drug effects; D002417:Cattle; D002453:Cell Cycle / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D006133:Growth Substances / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D007399:Interphase / Q000187:drug effects; D007700:Kinetics; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism*; D012501:Saphenous Vein / Q000166:cytology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D006133:Growth Substances; D006493:Heparin; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate