Decrease in growth factor receptors after treatment with serine protease inhibitor ONO-3403.
FOY-305 is a synthetic serine protease inhibitor and ONO-3403 is a derivative with a higher protease-inhibitory activity. The growth-suppressive effects of ONO-3403 were more prominent in Ha-ras-transformed NIH3T3 (ras-NIH) cells than in non-transformed NIH3T3 cells. After treatment of ras-NIH cells with ONO-3403 at 100-200 microg/ml, the percentage of cells found in G(1) phase decreased and, concomitantly, that in S phase increased. Molecular events caused by ONO-3403 were investigated by Western blot analysis using anti-phosphotyrosine antibody. The results showed a marked decrease in the tyrosine phosphorylation level of a 180-kDa protein after treatment with ONO-3403. This 180-kDa phosphotyrosine-containing molecule which was tentatively designated pY-p180 might be platelet-derived growth factor (PDGF) receptor since addition of PDGF to serum-starved NIH3T3 cells induced a marked tyrosine phosphorylation of the same size within 5 min. This was further confirmed by immunoprecipitation of cell extract with anti-PDGF-receptor antibody followed by Western blot analysis using anti-phosphotyrosine antibody. Treatment of T.Tn human esophageal carcinoma cells with ONO-3403 caused also decrease in pY-p180, which appeared to be epidermal growth factor receptor. Thus, ONO-3403 may induce growth suppression by down-regulation of cell surface growth factor receptors.
Hiwasa
Takaki
T
Department of Biochemistry and Genetics, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan. hiwasa@med.m.chiba-u.ac.jp
Shimada
Hideaki
H
Ochiai
Takenori
T
Liu
Tian-Ling
TL
Yuasa
Yasuhito
Y
Takiguchi
Masaki
M
Ohkoshi
Motohiro
M
D016475:3T3 Cells / Q000187:drug effects* / Q000378:metabolism; D000501:Allylglycine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000818:Animals; D001550:Benzamidines / Q000494:pharmacology*; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D066246:ErbB Receptors / Q000378:metabolism*; D005434:Flow Cytometry; D005562:Formazans; D051379:Mice; D010766:Phosphorylation; D011233:Precipitin Tests; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013778:Tetrazolium Salts; D014443:Tyrosine / Q000378:metabolism
D001550:Benzamidines; D005562:Formazans; D015842:Serine Proteinase Inhibitors; D013778:Tetrazolium Salts; C100807:ethyl N-allyl-N-(2-methyl-3-(4-(4-amidinophenoxycarbonyl)phenyl)propenoyl)aminoacetate methanesulfonate; D000501:Allylglycine; C079782:MTT formazan; D014443:Tyrosine; D066246:ErbB Receptors; D017479:Receptors, Platelet-Derived Growth Factor