title: Actinomycin D identified as an inhibitor of discoidin domain receptor 2 interaction with collagen through an insect cell based screening of a drug compound library.
abstract: Discoidin domain receptors belong to the cell surface receptor tyrosine kinase family and recognize collagens for their activating ligands. They have been implicated for cell growth and migration and their elevated expressions were observed in various human cancers. When we expressed human Discoidin domain receptor 2 (DDR2) in insect cells, the protein was targeted properly into the cell membrane and this could enforce the cells to adhere on culture plate coated with type I collagen. By taking advantage of this, we established a novel insect cell based screening protocol to identify chemicals which inhibit the interaction between DDR2 and collagen. We screened a drug-compound library to select an anti-cancer drug, actinomycin D, as the inhibitory compound. Actinomycin D prevented the activation of DDR2 by type I collagen in human embryonic kidney 293 cells with an IC(50) value of 9 microM, while it did not interfere with the activation of other receptor tyrosine kinases by their ligands. In conclusion we identified a new biological function of actinomycin D and the insect cell based method provides a useful protocol for screening inhibitors against the association of DDR2 with collagen.
authors.affiliation: Biomedical Research Center, Korea Institute of Science and Technology, Seoul.
authors.forename: Kauser
authors.initials: K
authors.lastname: Siddiqui
authors.forename: Geun Woong
authors.initials: GW
authors.lastname: Kim
authors.forename: Dong Hoon
authors.initials: DH
authors.lastname: Lee
authors.forename: Hae Ryun
authors.initials: HR
authors.lastname: Shin
authors.forename: Eun Gyeong
authors.initials: EG
authors.lastname: Yang
authors.forename: Nam Taek
authors.initials: NT
authors.lastname: Lee
authors.forename: Beom-Seok
authors.initials: BS
authors.lastname: Yang
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002461:Cell Line, Transformed; D024042:Collagen Type I / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology*; D000070818:Discoidin Domain Receptors; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D007313:Insecta; D019151:Peptide Library; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D024042:Collagen Type I; D019151:Peptide Library; D011975:Receptors, Mitogen; D011994:Recombinant Proteins; D014415:Tunicamycin; D003609:Dactinomycin; D000070818:Discoidin Domain Receptors; D020794:Receptor Protein-Tyrosine Kinases