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. 2014 Oct 27:(92):e51998.
doi: 10.3791/51998.

The soft agar colony formation assay

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

The soft agar colony formation assay

Stanley Borowicz et al. J Vis Exp. .

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.

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