title: Alternative to the soft-agar assay that permits high-throughput drug and genetic screens for cellular transformation.
abstract: Colony formation in soft agar is the gold-standard assay for cellular transformation in vitro, but it is unsuited for high-throughput screening. Here, we describe an assay for cellular transformation that involves growth in low attachment (GILA) conditions and is strongly correlated with the soft-agar assay. Using GILA, we describe high-throughput screens for drugs and genes that selectively inhibit or increase transformation, but not proliferation. Such molecules are unlikely to be found through conventional drug screening, and they include kinase inhibitors and drugs for noncancer diseases. In addition to known oncogenes, the genetic screen identifies genes that contribute to cellular transformation. Lastly, we demonstrate the ability of Food and Drug Administration-approved noncancer drugs to selectively kill ovarian cancer cells derived from patients with chemotherapy-resistant disease, suggesting this approach may provide useful information for personalized cancer treatment.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115;
authors.forename: Asaf
authors.initials: A
authors.lastname: Rotem
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115;
authors.forename: Andreas
authors.initials: A
authors.lastname: Janzer
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02115; and Broad Institute of Harvard and MIT, Cambridge, MA 02139.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Izar
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115;
authors.forename: Zhe
authors.initials: Z
authors.lastname: Ji
authors.affiliation: Broad Institute of Harvard and MIT, Cambridge, MA 02139.
authors.forename: John G
authors.initials: JG
authors.lastname: Doench
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02115; and Broad Institute of Harvard and MIT, Cambridge, MA 02139.
authors.forename: Levi A
authors.initials: LA
authors.lastname: Garraway
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115; kevin@hms.harvard.edu.
authors.forename: Kevin
authors.initials: K
authors.lastname: Struhl
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D000362:Agar / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000188:drug therapy / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D002471:Cell Transformation, Neoplastic; D002626:Chemistry, Pharmaceutical; D015195:Drug Design; D004354:Drug Screening Assays, Antitumor; D005260:Female; D005347:Fibroblasts / Q000473:pathology; D005434:Flow Cytometry; D057166:High-Throughput Screening Assays; D006801:Humans; D016366:Open Reading Frames; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000473:pathology
chemical_list: D000970:Antineoplastic Agents; D000255:Adenosine Triphosphate; D000362:Agar
keywords: cancer; drug screen; genetic screen; personalized medicine; transformation