Inhibition of cell growth: effects of the tyrosine kinase inhibitor CGP 53716.
The growth factors, platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF) play major roles in enhanced smooth muscle cells growth in rodent blood vessels after vascular injury. Tyrosine kinase inhibition has been shown to be effective in blocking tyrosine phosphorylation at the PDGF and bFGF receptors in cultured fibroblast and vascular smooth muscle cells which in turn inhibits their proliferation. Our study evaluated the PDGF selective tyrosine kinase inhibitor, CGP 53716, on serum, PDGF-BB, bFGF or epidermal growth factor-induced growth responses in cultured rat aortic smooth muscle cells (RASMC) and Balb/3T3 fibroblasts (3T3). CGP 53716 inhibited serum-induced cell growth in RASMC, but not in 3T3 cells. CGP 53716 completely blocked PDGF-BB tyrosine receptor autophosphorylation in RASMC and 3T3 cells, PDGF-BB-induced phosphorylation of mitogen-activated protein kinase at 1 microM in RASMC and inhibited PDGF-BB-induced c-Fos protein expression at 1 microM in RASMC; consistent with inhibition of PDGF-BB-induced DNA synthesis. To examine the selectivity of CGP 53716, PDGF-BB, bFGF or EGF-induced DNA synthesis was measured using thymidine incorporation. CGP 53716 inhibited PDGF-BB-, bFGF- and EGF-induced DNA synthesis in a concentration-dependent manner in each cell line. CGP 53716 showed a 2- to 4-fold selectivity for PDGF-BB-stimulated DNA synthesis over bFGF or EGF in RASMC or 3T3 cells. To rule out that bFGF induced the release of endogenous PDGF, an antibody to PDGF-AB, which binds to all three isoforms of PDGF, was coincubated with bFGF and did not suppress the DNA synthesis induced by bFGF. Based on these results, CGP 53716 is not selective for the PDGF receptor as previously reported. However, EGF-stimulated receptor autophosphorylation of mitogen-activated protein kinase phosphorylation and c-Fos protein expression were not inhibited by CGP 53716 at 1 or 10 microM in RASMC. These findings suggest that CGP 53716 may inhibit multiple growth factor pathways as indicated by inhibition of DNA synthesis. However, these effects must be downstream from the signaling for c-Fos protein expression or use an alternate signaling route. These results further suggest that CGP 53716 may have a therapeutic potential for the treatment of vascular proliferative diseases which are stimulated by not only PDGF but other growth factors such as bFGF and EGF.
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Department of Vascular and Cardiac Diseases, Parke-Davis Pharmaceutical Research Division, Warner Lambert Company, Ann Arbor, Michigan 48105, USA.
Keiser
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JA
D016475:3T3 Cells; D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006131:Growth Inhibitors / Q000494:pharmacology*; D051379:Mice; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011725:Pyridines / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D051381:Rats
C092972:CGP 53716; D004791:Enzyme Inhibitors; D006131:Growth Inhibitors; D010982:Platelet-Derived Growth Factor; D011725:Pyridines; D011743:Pyrimidines; D004815:Epidermal Growth Factor; D004247:DNA; D011505:Protein-Tyrosine Kinases