title: Protein-based nanotoxicology assessment strategy.
abstract: The nanomaterial community calls for standardized in vitro assays to determine nanoparticle toxicity in the effort to reduce the number of in vivo validation experiments. We demonstrate that chip-based protein detection is suitable for assessing toxicity and may complement traditional assays to improve selection of primary hits for subsequent analysis. As nanodrug mimics, we analyzed the effect of transiently transfected siRNAs in MCF7 breast cancer cells and normal MCF12A breast cells, resembling a differential screen. As a measure of cytotoxicity, we determined cell viability as well as protein expression of glyceraldehyde-3-phosphate dehydrogenase, transferrin receptor, and the proliferation marker Ki67. The evaluation of cell lethality and protein expression unraveled cellular effects overseen by one method alone.
authors.affiliation: Lundbeckfonden Center of Excellence in Nanomedicine NanoCAN, University of Southern Denmark, Odense, Denmark; Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark. Electronic address: mlpedersen@health.sdu.dk.
authors.forename: Marlene Pedersen
authors.initials: MP
authors.lastname: Elnegaard
authors.affiliation: Lundbeckfonden Center of Excellence in Nanomedicine NanoCAN, University of Southern Denmark, Odense, Denmark; Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark; Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.
authors.forename: Markus
authors.initials: M
authors.lastname: List
authors.affiliation: Lundbeckfonden Center of Excellence in Nanomedicine NanoCAN, University of Southern Denmark, Odense, Denmark; Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
authors.forename: Helle
authors.initials: H
authors.lastname: Christiansen
authors.affiliation: Lundbeckfonden Center of Excellence in Nanomedicine NanoCAN, University of Southern Denmark, Odense, Denmark; Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
authors.forename: Steffen
authors.initials: S
authors.lastname: Schmidt
authors.affiliation: Lundbeckfonden Center of Excellence in Nanomedicine NanoCAN, University of Southern Denmark, Odense, Denmark; Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
authors.forename: Jan
authors.initials: J
authors.lastname: Mollenhauer
authors.affiliation: Lundbeckfonden Center of Excellence in Nanomedicine NanoCAN, University of Southern Denmark, Odense, Denmark; Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
authors.forename: Ines
authors.initials: I
authors.lastname: Block
mesh_terms: D001940:Breast / Q000166:cytology; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006801:Humans; D019394:Ki-67 Antigen / Q000235:genetics; D053758:Nanoparticles / Q000633:toxicity*; D040081:Protein Array Analysis / Q000379:methods*; D034741:RNA, Small Interfering / Q000235:genetics; D011990:Receptors, Transferrin / Q000235:genetics; D018675:Toxicity Tests / Q000379:methods*; D014162:Transfection
chemical_list: D019394:Ki-67 Antigen; D034741:RNA, Small Interfering; D011990:Receptors, Transferrin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: Cell viability; Nanotoxicology; Protein array; RNAi