The Soft Agar Colony Formation Assay

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 string...

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Published in:Journal of visualized experiments no. 92; p. e51998
Main Authors: Borowicz, Stanley, Van Scoyk, Michelle, Avasarala, Sreedevi, Karuppusamy Rathinam, Manoj Kumar, Tauler, Jordi, Bikkavilli, Rama Kamesh, Winn, Robert A.
Format: Journal Article
Language:English
Published: United States MyJove Corporation 27.10.2014
Subjects:
agar
Animals
carcinogenesis
Cell Line, Tumor
cell lines
Cellular Biology
Colony-Forming Units Assay - methods
ligands
lung neoplasms
Lung Neoplasms - pathology
Mice
neoplasm cells
signal transduction
Tumor Stem Cell Assay - methods
ISSN:1940-087X, 1940-087X
Online Access:Get full text
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Summary: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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Correspondence to: Rama Kamesh Bikkavilli at kamesh@uic.edu
ISSN:1940-087X
1940-087X
DOI:10.3791/51998