软琼脂集落形成试验。
The soft agar colony formation assay.
作者信息
Borowicz Stanley, Van Scoyk Michelle, Avasarala Sreedevi, Karuppusamy Rathinam Manoj Kumar, Tauler Jordi, Bikkavilli Rama Kamesh, Winn Robert A
机构信息
Department of Hematology and Oncology, University of Illinois at Chicago.
Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago.
出版信息
J Vis Exp. 2014 Oct 27(92):e51998. doi: 10.3791/51998.
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.
不依赖贴壁生长是转化细胞在没有固体表面的情况下独立生长的能力，是致癌作用的一个标志。软琼脂集落形成试验是一种成熟的体外表征这种能力的方法，被认为是细胞恶性转化最严格的测试之一。该试验还允许对这种能力在各种处理条件下的反应进行半定量评估。在这里，我们将使用小鼠肺癌细胞系CMT167演示软琼脂集落形成试验，以证明Wnt信号通路的两个成员Wnt7A和卷曲蛋白9（Fzd-9）的肿瘤抑制作用。Wnt7a和Fzd-9的同时过表达导致CMT167细胞集落形成受到抑制。这表明Wnt7a配体及其卷曲蛋白9受体的表达足以抑制小鼠肺癌模型中的肿瘤生长。
Zotero 插件