A novel asymmetric 3D in-vitro assay for the study of tumor cell invasion.
A novel asymmetric 3D in-vitro assay for the study of tumor cell invasion.
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DOI：
10.1186/1471-2407-9-415
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发表时间：
2009-11-30
期刊：
影响因子：
3.8
通讯作者：
Neufeld G
中科院分区：
文献类型：
--
作者：
Brekhman V;Neufeld G
诱导肿瘤细胞侵袭是肿瘤发展的重要步骤。由于体内侵袭试验的成本和速度较慢，因此需要定量的体外侵袭试验，尽可能地模拟肿瘤环境，并严格控制条件。我们通过在两层胶原蛋白之间嵌入一层肿瘤细胞，建立了一种新的非对称3D体外侵袭试验。然后让细胞侵入胶原蛋白的上层和下层。为了可视化入侵细胞，将凝胶垂直于单层切片，以便在播种后单层呈现为细线，精确地定义入侵的起源。由此可以定量测定浸润肿瘤细胞的数量、增殖速率、浸润距离和浸润方向。该实验用于比较几种肿瘤细胞类型的侵袭性，结果与先前描述的实验结果相比较。赖氨酸氧化酶样蛋白-2 （Loxl2）是一种有效的侵袭诱导剂。通过我们的实验，我们首次发现抑制几种类型肿瘤细胞中的内源性Loxl2表达可以强烈抑制其侵袭性。我们还利用了该试验的不对称性质，以表明纤维连接蛋白比层粘连蛋白更有效地增强乳腺癌细胞的侵袭性。该实验的不对称特性也被用来证明来自成纤维细胞的可溶性因子可以优先吸引入侵的乳腺癌细胞。我们的检测方法比以前的侵袭检测方法有几个优势，因为它可以在模拟肿瘤微环境的3D环境中定量分析肿瘤细胞的定向侵袭行为。这对于研究肿瘤微环境成分对肿瘤细胞侵袭性的影响尤其有用。
The induction of tumor cell invasion is an important step in tumor progression. Due to the cost and slowness of in-vivo invasion assays, there is need for quantitative in-vitro invasion assays that mimic as closely as possible the tumor environment and in which conditions can be rigorously controlled. We have established a novel asymmetric 3D in-vitro invasion assay by embedding a monolayer of tumor cells between two layers of collagen. The cells were then allowed to invade the upper and lower layers of collagen. To visualize invading cells the gels were sectioned perpendicular to the monolayer so that after seeding the monolayer appears as a thin line precisely defining the origin of invasion. The number of invading tumor cells, their proliferation rate, the distance they traverse and the direction of invasion could then be determined quantitatively. The assay was used to compare the invasive properties of several tumor cell types and the results compare well with those obtained by previously described assays. Lysyl-oxidase like protein-2 (Loxl2) is a potent inducer of invasiveness. Using our assay we show for the first time that inhibition of endogenous Loxl2 expression in several types of tumor cells strongly inhibits their invasiveness. We also took advantage of the asymmetric nature of the assay in order to show that fibronectin enhances the invasiveness of breast cancer cells more potently than laminin. The asymmetric properties of the assay were also used to demonstrate that soluble factors derived from fibroblasts can preferentially attract invading breast cancer cells. Our assay displays several advantages over previous invasion assays as it is allows the quantitative analysis of directional invasive behavior of tumor cells in a 3D environment mimicking the tumor microenvironment. It should be particularly useful for the study of the effects of components of the tumor microenvironment on tumor cell invasiveness.
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影响因子：
11.2
作者：
Studebaker, Adam W.;Storci, Gianluca;Hall, Brett M.
通讯作者：
Hall, Brett M.
DOI：
10.1083/jcb.200209006
发表时间：
2003-01-20
期刊：
The Journal of cell biology
影响因子：
--
作者：
Wolf K;Mazo I;Leung H;Engelke K;von Andrian UH;Deryugina EI;Strongin AY;Bröcker EB;Friedl P
通讯作者：
Friedl P
影响因子：
--
作者：
Kleinman, H K;Jacob, K
通讯作者：
Jacob, K
影响因子：
11.4
作者：
Peinado, H;Iglesias-de la Cruz, MD;Portillo, F
通讯作者：
Portillo, F
影响因子：
3.5
作者：
Maquoi, E;Noël, A;Foidart, JM
通讯作者：
Foidart, JM