title: Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells.
abstract: Given the inherent difficulties in investigating the mechanisms of tumor progression in vivo, cell-based assays such as the soft agar colony formation assay (hereafter called soft agar assay), which measures the ability of cells to proliferate in semi-solid matrices, remain a hallmark of cancer research. A key advantage of this technique over conventional 2D monolayer or 3D spheroid cell culture assays is the close mimicry of the 3D cellular environment to that seen in vivo. Importantly, the soft agar assay also provides an ideal tool to rigorously test the effects of novel compounds or treatment conditions on cell proliferation and migration. Additionally, this assay enables the quantitative assessment of cell transformation potential within the context of genetic perturbations. We recently identified peptidylarginine deiminase 2 (PADI2) as a potential breast cancer biomarker and therapeutic target. Here we highlight the utility of the soft agar assay for preclinical anti-cancer studies by testing the effects of the PADI inhibitor, BB-Cl-amidine (BB-CLA), on the tumorigenicity of human ductal carcinoma in situ (MCF10DCIS) cells.
authors.affiliation: Department of Baker Institute for Animal Health, Cornell University.
authors.forename: Sachi
authors.initials: S
authors.lastname: Horibata
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University.
authors.forename: Tommy V
authors.initials: TV
authors.lastname: Vo
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School.
authors.forename: Venkataraman
authors.initials: V
authors.lastname: Subramanian
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Thompson
authors.affiliation: Department of Baker Institute for Animal Health, Cornell University; sac269@cornell.edu.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Coonrod
mesh_terms: D000362:Agar; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology*; D063646:Carcinogenesis / Q000187:drug effects; D002278:Carcinoma in Situ / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D018450:Disease Progression; D004354:Drug Screening Assays, Antitumor; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D006867:Hydrolases / Q000037:antagonists & inhibitors; D014411:Neoplastic Stem Cells / Q000187:drug effects*; D009952:Ornithine / Q000031:analogs & derivatives / Q000494:pharmacology; D000080064:Protein-Arginine Deiminase Type 2; D000076342:Protein-Arginine Deiminases; D012685:Sepharose / Q000737:chemistry
chemical_list: D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; C558727:N-alpha-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine amide; D000362:Agar; D012685:Sepharose; D009952:Ornithine; D006867:Hydrolases; C000620352:PADI2 protein, human; D000080064:Protein-Arginine Deiminase Type 2; D000076342:Protein-Arginine Deiminases