title: The soft agar colony formation assay.
abstract: Anchorage-independent growth is the ability of transformed cells to grow independently of a solid surface, and is a hallmark of carcinogenesis. The soft agar colony formation assay is a well-established method for characterizing this capability in vitro and is considered to be one of the most stringent tests for malignant transformation in cells. This assay also allows for semi-quantitative evaluation of this capability in response to various treatment conditions. Here, we will demonstrate the soft agar colony formation assay using a murine lung carcinoma cell line, CMT167, to demonstrate the tumor suppressive effects of two members of the Wnt signaling pathway, Wnt7A and Frizzled-9 (Fzd-9). Concurrent overexpression of Wnt7a and Fzd-9 caused an inhibition of colony formation in CMT167 cells. This shows that expression of Wnt7a ligand and its Frizzled-9 receptor is sufficient to suppress tumor growth in a murine lung carcinoma model.
authors.affiliation: Department of Hematology and Oncology, University of Illinois at Chicago.
authors.forename: Stanley
authors.initials: S
authors.lastname: Borowicz
authors.affiliation: Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago.
authors.forename: Michelle
authors.initials: M
authors.lastname: Van Scoyk
authors.affiliation: Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago.
authors.forename: Sreedevi
authors.initials: S
authors.lastname: Avasarala
authors.affiliation: Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago.
authors.forename: Manoj Kumar
authors.initials: MK
authors.lastname: Karuppusamy Rathinam
authors.affiliation: Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago.
authors.forename: Jordi
authors.initials: J
authors.lastname: Tauler
authors.affiliation: Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago; kamesh@uic.edu.
authors.forename: Rama Kamesh
authors.initials: RK
authors.lastname: Bikkavilli
authors.affiliation: Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago; Jesse Brown Veterans Affairs Medical Center.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Winn
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D003114:Colony-Forming Units Assay / Q000379:methods*; D008175:Lung Neoplasms / Q000473:pathology; D051379:Mice; D014410:Tumor Stem Cell Assay / Q000379:methods