The Soft Agar Colony Formation Assay
The Soft Agar Colony Formation Assay
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DOI：
10.3791/51998
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发表时间：
2014-10-01
影响因子：
1.2
通讯作者：
Winn, Robert A.
中科院分区：
文献类型：
--
作者：
Borowicz, Stanley;Van Scoyk, Michelle;Winn, Robert A.
锚定非依赖性生长是转化细胞独立于固体表面生长的能力，并且是癌发生的标志。软琼脂集落形成试验是体外表征这种能力的成熟方法，被认为是细胞恶性转化的最严格试验之一。该测定还允许响应于各种处理条件的这种能力的半定量评价。在这里，我们将使用鼠肺癌细胞系CMT 167证明软琼脂集落形成试验，以证明Wnt信号通路的两个成员Wnt 7A和Frizzled-9（Fzd-9）的肿瘤抑制作用。Wnt 7a和Fzd-9的同时过表达引起CMT 167细胞中集落形成的抑制。这表明Wnt 7a配体及其Frizzled-9受体的表达足以抑制鼠肺癌模型中的肿瘤生长。
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.