Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells
Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells
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DOI：
10.3791/52727
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发表时间：
2015-05-01
影响因子：
1.2
通讯作者：
Coonrod, Scott A.
中科院分区：
文献类型：
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作者：
Horibata, Sachi;Vo, Tommy V.;Coonrod, Scott A.
考虑到在体内研究肿瘤进展机制的固有困难，基于细胞的实验，如软琼脂集落形成实验（以下称为软琼脂实验），测量细胞在半固体基质中的增殖能力，仍然是癌症研究的一个标志。与传统的2D单层或3D球形细胞培养分析相比，该技术的一个关键优势是3D细胞环境与体内观察到的密切模仿。重要的是，软琼脂实验也提供了一个理想的工具来严格测试新化合物或处理条件对细胞增殖和迁移的影响。此外，该分析能够在遗传扰动的背景下对细胞转化潜力进行定量评估。我们最近发现肽精氨酸脱亚胺酶2 （PADI2）是一种潜在的乳腺癌生物标志物和治疗靶点。在这里，我们通过测试PADI抑制剂BB-Cl-amidine （BB-CLA）对人导管原位癌（MCF10DCIS）细胞致瘤性的影响，强调软琼脂试验在临床前抗癌研究中的应用。
Given the inherent difficulties in investigating the mechanisms of tumor progression in vivo, cell-based assays such as the soft agar colony formation assay (hereafter called soft agar assay), which measures the ability of cells to proliferate in semi-solid matrices, remain a hallmark of cancer research. A key advantage of this technique over conventional 2D monolayer or 3D spheroid cell culture assays is the close mimicry of the 3D cellular environment to that seen in vivo. Importantly, the soft agar assay also provides an ideal tool to rigorously test the effects of novel compounds or treatment conditions on cell proliferation and migration. Additionally, this assay enables the quantitative assessment of cell transformation potential within the context of genetic perturbations. We recently identified peptidylarginine deiminase 2 (PADI2) as a potential breast cancer biomarker and therapeutic target. Here we highlight the utility of the soft agar assay for preclinical anti-cancer studies by testing the effects of the PADI inhibitor, BB-Cl-amidine (BB-CLA), on the tumorigenicity of human ductal carcinoma in situ (MCF10DCIS) cells.