title: Nitroaspirin (NCX-4016), an NO donor, is antiangiogenic through induction of loss of redox-dependent viability and cytoskeletal reorganization in endothelial cells.
abstract: We recently reported that NCX-4016, a derivative of aspirin containing a nitro moiety that releases nitric oxide (NO) in a sustained fashion in biologic systems, is a potent cytotoxic agent inhibiting the proliferation of cisplatin-resistant human ovarian cancer cells. Therefore, we hypothesize that NCX-4016 possesses antiangiogenic properties. Our study with the bovine lung microvascular endothelial cells (BLMVECs) revealed that NCX-4016 significantly induced the loss of redox-dependent cell viability in a dose- and time-dependent manner, as assayed by the redox-sensitive Alamar blue cell viability assay. Fluorescence microscopy of cells labeled with NO-specific fluorophore (DAF-FM) confirmed that NCX-4016 generated significant levels of intracellular NO. NO donors, including S-nitroso-N-acetylpenicillamine, spermine NONOate, and isosorbide dinitrite, were less effective in causing loss of cell viability. Thiol-protectant, N-acetylcysteine, significantly attenuated the NCX-4016-induced loss of cell viability, suggesting the role of alteration of thiol-redox status therein. NCX-4016 also suppressed oxygen consumption, decreased transendothelial electrical resistance (EC barrier dysfunction), and induced actin cytoskeletal reorganization in BLMVECs. The in vitro assay with human umbilical vein ECs and BLMVECs revealed that NCX-4016, in a dose-dependent manner, significantly inhibited angiogenesis with almost complete inhibition at a 100-microM concentration, suggesting that NCX-4016 can act as an antiangiogenic drug.
authors.affiliation: Department of Internal Medicine, Divisions of Cardiology and Pulmonary, Critical Care, and Sleep Medicine, Dorothy M. Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University, Columbus, Ohio 43210, USA. kuppusamy.1@osu.edu
authors.forename: Narasimham L
authors.initials: NL
authors.lastname: Parinandi
authors.forename: Ashish
authors.initials: A
authors.lastname: Sharma
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Eubank
authors.forename: Bruce F
authors.initials: BF
authors.lastname: Kaufman
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Kutala
authors.forename: Clay B
authors.initials: CB
authors.lastname: Marsh
authors.forename: Louis J
authors.initials: LJ
authors.lastname: Ignarro
authors.forename: Periannan
authors.initials: P
authors.lastname: Kuppusamy
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001241:Aspirin / Q000031:analogs & derivatives* / Q000494:pharmacology; D002196:Capillaries / Q000166:cytology; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008168:Lung / Q000098:blood supply; D008856:Microscopy, Fluorescence; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D010084:Oxidation-Reduction; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D000975:Antioxidants; D020030:Nitric Oxide Donors; D006861:Hydrogen Peroxide; C102148:nitroaspirin; D001241:Aspirin