title: Development of a high-throughput three-dimensional invasion assay for anti-cancer drug discovery.
abstract: The lack of three-dimensional (3-D) high-throughput (HT) screening assays designed to identify anti-cancer invasion drugs is a major hurdle in reducing cancer-related mortality, with the key challenge being assay standardization. Presented is the development of a novel 3-D invasion assay with HT potential that involves surrounding cell-collagen spheres within collagen to create a 3-D environment through which cells can invade. Standardization was achieved by designing a tooled 96-well plate to create a precisely designated location for the cell-collagen spheres and by using dialdehyde dextran to inhibit collagen contraction, maintaining uniform size and shape. This permits automated readout for determination of the effect of inhibitory compounds on cancer cell invasion. Sensitivity was demonstrated by the ability to distinguish varying levels of invasiveness of cancer cell lines, and robustness was determined by calculating the Z-factor. A Z-factor of 0.65 was obtained by comparing the effects of DMSO and anti-β1-integrin antibody, an inhibitory reagent, on the invasion of Du145 cancer cells, suggesting this novel assay is suitable for large scale drug discovery. As proof of principle, the NCI Diversity Compound Library was screened against human invasive cancer cells. Nine compounds exhibiting high potency and low toxicity were identified, including DX-52-1, a compound previously reported to inhibit cell migration, a critical determinant of cancer invasion. The results indicate that this innovative HT platform is a simple, precise, and easy to replicate 3-D invasion assay for anti-cancer drug discovery.
authors.affiliation: Department of Medicine/Cancer Prevention, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Nikki A
authors.initials: NA
authors.lastname: Evensen
authors.affiliation: Department of Medicine/Cancer Prevention, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Preventive Medicine, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Tissue Engineering, Stevens Institute of Technology, Hoboken, New Jersey, United States of America.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Chemistry, Stony Brook University, Stony Brook, New York, United States of America , and.
authors.forename: Nicole S
authors.initials: NS
authors.lastname: Sampson
authors.affiliation: Department of Research, Veterans Affair Medical Center, Northport, New York, United States of America.
authors.forename: Stanley
authors.initials: S
authors.lastname: Zucker
authors.affiliation: Department of Medicine/Cancer Prevention, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Jian
authors.initials: J
authors.lastname: Cao
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology / Q000633:toxicity; D045744:Cell Line, Tumor; D055808:Drug Discovery; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans; D009361:Neoplasm Invasiveness; D015203:Reproducibility of Results; D018874:Spheroids, Cellular; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents