The soft agar colony formation assay

  • J Vis Exp. 2014 Oct 27:(92):e51998. doi: 10.3791/51998.
Stanley Borowicz  1 ,  Michelle Van Scoyk  2 ,  Sreedevi Avasarala  2 ,  Manoj Kumar Karuppusamy Rathinam  2 ,  Jordi Tauler  2 ,  Rama Kamesh Bikkavilli  3 ,  Robert A Winn  4
Affiliations
  • 1. Department of Hematology and Oncology, University of Illinois at Chicago.
  • 2. Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago.
  • 3. Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago; [email protected].
  • 4. Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago; Jesse Brown Veterans Affairs Medical Center.
PMID: 25408172 DOI: 10.3791/51998
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.