title: The first EGF domain of coagulation factor IX attenuates cell adhesion and induces apoptosis.
abstract: Coagulation factor IX (FIX) is an essential plasma protein for blood coagulation. The first epidermal growth factor (EGF) motif of FIX (EGF-F9) has been reported to attenuate cell adhesion to the extracellular matrix (ECM). The purpose of the present study was to determine the effects of this motif on cell adhesion and apoptosis. Treatment with a recombinant EGF-F9 attenuated cell adhesion to the ECM within 10 min. De-adhesion assays with native FIX recombinant FIX deletion mutant proteins suggested that the de-adhesion activity of EGF-F9 requires the same process of FIX activation as that which occurs for coagulation activity. The recombinant EGF-F9 increased lactate dehydrogenase (LDH) activity release into the medium and increased the number of cells stained with annexin V and activated caspase-3, by 8.8- and 2.7-fold respectively, indicating that EGF-F9 induced apoptosis. Activated caspase-3 increased very rapidly after only 5 min of administration of recombinant EGF-F9. Treatment with EGF-F9 increased the level of phosphorylated p38 mitogen-activated protein kinase (MAPK), but not that of phosphorylated MAPK 44/42 or c-Jun N-terminal kinase (JNK). Inhibitors of caspase-3 suppressed the release of LDH. Caspase-3 inhibitors also suppressed the attenuation of cell adhesion and phosphorylation of p38 MAPK by EGF-F9. Our data indicated that EGF-F9 activated signals for apoptosis and induced de-adhesion in a caspase-3 dependent manner.
authors.affiliation: Division of Physiology, Department of Biomedical Science, School of Medicine, Nihon University, 30-1, Oyaguchi Kami-cho, Itabashi-ku, Tokyo 173-8610, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Ishikawa
authors.affiliation: Division of Dental Surgery, School of Medicine, Nihon University, 30-1, Oyaguchi Kami-cho, Itabashi-ku, Tokyo 173-8610, Japan.
authors.forename: Hisataka
authors.initials: H
authors.lastname: Kitano
authors.affiliation: Division of Dental Surgery, School of Medicine, Nihon University, 30-1, Oyaguchi Kami-cho, Itabashi-ku, Tokyo 173-8610, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Mamiya
authors.affiliation: Division of Physiology, Department of Biomedical Science, School of Medicine, Nihon University, 30-1, Oyaguchi Kami-cho, Itabashi-ku, Tokyo 173-8610, Japan.
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Kokubun
authors.affiliation: Division of Physiology, Department of Biomedical Science, School of Medicine, Nihon University, 30-1, Oyaguchi Kami-cho, Itabashi-ku, Tokyo 173-8610, Japan hidai.chiaki@nihon-u.ac.jp.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Hidai
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D023102:Anoikis / Q000235:genetics; D017209:Apoptosis / Q000235:genetics*; D001777:Blood Coagulation / Q000235:genetics*; D053148:Caspase 3 / Q000235:genetics* / Q000378:metabolism; D002448:Cell Adhesion; D004789:Enzyme Activation / Q000235:genetics; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism; D005164:Factor IX / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000235:genetics; D017384:Sequence Deletion; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D005164:Factor IX; D007770:L-Lactate Dehydrogenase; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: adhesion; anoikis; apoptosis; epidermal growth factor; factor IX