Stimulation of heparan sulfate proteoglycan synthesis and secretion during G1 phase induced by growth factors and PMA.
Fetal calf serum (FCS) and PMA (phorbol 12-myristate-13-acetate) specifically stimulate the synthesis of heparan sulfate proteoglycan in endothelial cells. Staurosporine and n-butanol, kinase inhibitors, abolish the PMA effect. Forskolin and 8-bromo adenosine 3':5'-cyclic monophosphate, activators of, respectively, adenylate cyclase and protein kinase A cannot reproduce the PMA effect. The kinetics of cell entry into S phase of the endothelial cells was determined by DNA synthesis ([3H]-thymidine and Br-dU incorporation), and flow cytometry. The mitogenic effect of fetal calf serum is abolished by PMA. Also, PMA pre-treatment inhibits the enhanced synthesis of heparan sulfate proteoglycan after a second PMA exposure. Remarkably, the stimulation of heparan sulfate proteoglycan synthesis by fetal calf serum and PMA seems to be mainly restricted to G1 phase. Therefore fetal calf serum and PMA cause an enhanced synthesis of heparan sulfate proteoglycan, and PMA causes a cell cycle block at G1 phase.
Departamento de Bioquímica, Escola Paulista de Medicina, UNIFESP, São Paulo, SP, Brazil.
M A
MA
Porcionatto
C R
CR
Moreira
C F
CF
Lotfi
H A
HA
Armelin
C P
CP
Dietrich
H B
HB
Nader
D000818:Animals; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D004789:Enzyme Activation; D005434:Flow Cytometry; D016193:G1 Phase; D006133:Growth Substances / Q000494:pharmacology*; D019812:Heparan Sulfate Proteoglycans / Q000096:biosynthesis* / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D011817:Rabbits; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D006133:Growth Substances; D019812:Heparan Sulfate Proteoglycans; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate