The Soft Agar Colony Formation Assay

被引：468

作者：

Borowicz, Stanley ^{[1
]}

Van Scoyk, Michelle ^{[2
]}

Avasarala, Sreedevi ^{[2
]}

Rathinam, Manoj Kumar Karuppusamy ^{[2
]}

Tauler, Jordi ^{[2
]}

Bikkavilli, Rama Kamesh ^{[2
]}

Winn, Robert A. ^{[2
,3
]}

机构：

[1] Univ Illinois, Dept Hematol & Oncol, Chicago, IL USA

[2] Univ Illinois, Dept Pulm Crit Care Sleep & Allergy, Chicago, IL 60607 USA

[3] Jesse Brown Vet Affairs Med Ctr, Chicago, IL USA

来源：

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2014年 / 92期

关键词：

Cellular Biology; Issue; 92; Wnt; Frizzled; Soft Agar Assay; Colony Formation Assay; tumor suppressor; lung cancer; CELL LUNG-CANCER; STEM-CELLS; GROWTH;

D O I：

10.3791/51998

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：6

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