Dynamic and influential interaction of cancer cells with normal epithelial cells in 3D culture.
Dynamic and influential interaction of cancer cells with normal epithelial cells in 3D culture.
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DOI：
10.1186/s12935-014-0108-6
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发表时间：
2014
影响因子：
5.8
通讯作者：
Itasaki N
中科院分区：
文献类型：
--
作者：
Ivers LP;Cummings B;Owolabi F;Welzel K;Klinger R;Saitoh S;O'Connor D;Fujita Y;Scholz D;Itasaki N
癌症微环境对癌细胞的生长和动力学有很大影响。然而，传统的2D培养系统不能反映体内条件，阻碍了对癌细胞动力学的详细研究。这项工作的目的是建立一种方法来揭示癌症和正常上皮细胞的相互作用，使用3D时间推移。将GFP标记的乳腺癌细胞MDA-MB-231与mCherry标记的非癌上皮细胞MDCK在凝胶基质中共培养。在3D培养中，上皮细胞建立球形形态（上皮球），从而为癌细胞提供上皮基底表面的可接近性，类似于体内条件。使用延时分析监测细胞运动。超微结构，免疫细胞化学和蛋白质表达分析也进行了延时研究。在2D培养系统中，大多数MDA-MB-231细胞作为单细胞表现出梭形形态，与此相反，在3D培养中，发现MDA-MB-231细胞是单细胞或形成聚集体，两者都是能动的。单个MDA-MB-231细胞表现出圆形和纺锤形，从一种形状到另一种形状的动态变化在几个小时内可见。当与上皮细胞共培养时，MDA-MB-231细胞表现出对上皮球的强烈吸引，并继续包围和吞噬上皮细胞团。被包围的上皮细胞最终被破坏，成为碎片，并被带入MDA-MB-231细胞。然而，当存在相对大的正常上皮细胞群体时，尽管反复接触，MDA-MB-231细胞也不能有效地吞噬上皮球。与大量正常上皮细胞共培养的MDA-MB-231细胞显示单羧酸转运蛋白-1的表达减少，表明细胞代谢发生变化。还观察到明胶消化能力水平降低以及基质金属蛋白酶-2的产生减少。这种培养方法是研究癌细胞动力学和响应微环境的细胞变化的有力技术。所述方法可用于多个方面，例如：癌症和非癌症细胞类型的不同组合，解决癌细胞对宿主细胞的器官特异性亲和力，以及监测对抗癌药物的细胞应答。本文的在线版本（doi：10.1186/s12935-014-0108-6）包含补充材料，可供授权用户使用。
The cancer microenvironment has a strong impact on the growth and dynamics of cancer cells. Conventional 2D culture systems, however, do not reflect in vivo conditions, impeding detailed studies of cancer cell dynamics. This work aims to establish a method to reveal the interaction of cancer and normal epithelial cells using 3D time-lapse. GFP-labelled breast cancer cells, MDA-MB-231, were co-cultured with mCherry-labelled non-cancerous epithelial cells, MDCK, in a gel matrix. In the 3D culture, the epithelial cells establish a spherical morphology (epithelial sphere) thus providing cancer cells with accessibility to the basal surface of epithelia, similar to the in vivo condition. Cell movement was monitored using time-lapse analyses. Ultrastructural, immunocytochemical and protein expression analyses were also performed following the time-lapse study. In contrast to the 2D culture system, whereby most MDA-MB-231 cells exhibit spindle-shaped morphology as single cells, in the 3D culture the MDA-MB-231 cells were found to be single cells or else formed aggregates, both of which were motile. The single MDA-MB-231 cells exhibited both round and spindle shapes, with dynamic changes from one shape to the other, visible within a matter of hours. When co-cultured with epithelial cells, the MDA-MB-231 cells displayed a strong attraction to the epithelial spheres, and proceeded to surround and engulf the epithelial cell mass. The surrounded epithelial cells were eventually destroyed, becoming debris, and were taken into the MDA-MB-231 cells. However, when there was a relatively large population of normal epithelial cells, the MDA-MB-231 cells did not engulf the epithelial spheres effectively, despite repeated contacts. MDA-MB-231 cells co-cultured with a large number of normal epithelial cells showed reduced expression of monocarboxylate transporter-1, suggesting a change in the cell metabolism. A decreased level of gelatin-digesting ability as well as reduced production of matrix metaroproteinase-2 was also observed. This culture method is a powerful technique to investigate cancer cell dynamics and cellular changes in response to the microenvironment. The method can be useful for various aspects such as; different combinations of cancer and non-cancer cell types, addressing the organ-specific affinity of cancer cells to host cells, and monitoring the cellular response to anti-cancer drugs. The online version of this article (doi:10.1186/s12935-014-0108-6) contains supplementary material, which is available to authorized users.
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影响因子：
4.8
作者：
Debnath, J;Muthuswamy, SK;Brugge, JS
通讯作者：
Brugge, JS
影响因子：
5.2
作者：
Lehmann, Christian;Jobs, Gabriele;Kubbies, Manfred
通讯作者：
Kubbies, Manfred
影响因子：
64.5
作者：
Hotary, KB;Allen, ED;Weiss, SJ
通讯作者：
Weiss, SJ
影响因子：
4.8
作者：
Mendez, Melissa G.;Kojima, Shin-Ichiro;Goldman, Robert D.
通讯作者：
Goldman, Robert D.
影响因子：
5.6
作者：
Hegedus, Luca;Cho, Hyojin;Eliceiri, George L.
通讯作者：
Eliceiri, George L.