Epithelial-mesenchymal transition stimulates human cancer cells to extend microtubule-based invasive protrusions and suppresses cell growth in collagen gel.
Epithelial-mesenchymal transition stimulates human cancer cells to extend microtubule-based invasive protrusions and suppresses cell growth in collagen gel.
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DOI：
10.1371/journal.pone.0053209
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Miyazaki K
中科院分区：
文献类型：
--
作者：
Oyanagi J;Ogawa T;Sato H;Higashi S;Miyazaki K
上皮间质转化（EMT）是肿瘤侵袭和转移的关键事件。然而，过去大多数 EMT 研究都是在传统的二维 (2D) 单层培养中进行的。因此，EMT 诱导的癌细胞获得何种侵袭性表型仍不清楚。为了解决这一点，我们尝试在更生理的三维 (3D) 胶原凝胶培养物中表征 EMT 细胞。通过用 TGF-β 处理三种人类癌细胞系（A549、Panc-1 和 MKN-1）来诱导 EMT。除了诱导众所周知的形态变化和 EMT 标记物表达外，TGF-β 处理还刺激这些细胞在 2D 培养物中过表达侵袭标记物层粘连蛋白 γ2 和 MT1-MMP。 EMT 诱导增强了 2D 培养中的细胞运动性以及对纤连蛋白和胶原蛋白的粘附性。尽管 EMT 细胞在 2D 培养中表现出与对照细胞相当的细胞生长，但它们的生长速率在软琼脂和胶原凝胶培养中受到极大抑制。最典型的是，EMT 诱导的癌细胞通常在胶原蛋白凝胶中显着扩展侵入性突起。这些突起主要由微管而不是肌动蛋白细胞骨架支撑。蜗牛引入的稳定 EMT 细胞在没有 TGF-ß 的 3D 条件下显示出类似的突起。此外，这些突起被秋水仙碱或热休克蛋白 90 (HSP-90) 和蛋白磷酸酶 2A 抑制剂抑制。然而，MMP 抑制剂并不能抑制突起的形成。这些数据表明，EMT 通过延伸微管突起和抑制细胞生长来增强肿瘤细胞浸润间质。细胞对纤连蛋白和胶原蛋白的粘附增强以及高细胞运动性似乎对于肿瘤侵袭也很重要。
Epithelial-mesenchymal transition (EMT) is a crucial event in tumor invasion and metastasis. However, most of past EMT studies have been conducted in the conventional two-dimensional (2D) monolayer culture. Therefore, it remains unclear what invasive phenotypes are acquired by EMT-induced cancer cells. To address this point, we attempted to characterize EMT cells in more physiological, three-dimensional (3D) collagen gel culture. EMT was induced by treating three human carcinoma cell lines (A549, Panc-1 and MKN-1) with TGF-ß. The TGF-ß treatment stimulated these cells to overexpress the invasion markers laminin γ2 and MT1-MMP in 2D culture, in addition to the induction of well-known morphological change and EMT marker expression. EMT induction enhanced cell motility and adhesiveness to fibronectin and collagen in 2D culture. Although EMT cells showed comparable cell growth to control cells in 2D culture, their growth rates were extremely suppressed in soft agar and collagen gel cultures. Most characteristically, EMT-induced cancer cells commonly and markedly extended invasive protrusions in collagen gel. These protrusions were mainly supported by microtubules rather than actin cytoskeleton. Snail-introduced, stable EMT cells showed similar protrusions in 3D conditions without TGF-ß. Moreover, these protrusions were suppressed by colchicine or inhibitors of heat shock protein 90 (HSP-90) and protein phosphatase 2A. However, MMP inhibitors did not suppress the protrusion formation. These data suggest that EMT enhances tumor cell infiltration into interstitial stroma by extending microtubule-based protrusions and suppressing cell growth. The elevated cell adhesion to fibronectin and collagen and high cell motility also seem important for the tumor invasion.
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DOI：
10.1038/nrm3141
发表时间：
2011-06-23
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
9.2
作者：
Gierke, Sarah;Wittmann, Torsten
通讯作者：
Wittmann, Torsten
影响因子：
4
作者：
Gawden-Bone, Christian;Zhou, Zhongjun;Lucocq, John
通讯作者：
Lucocq, John
影响因子：
2.3
作者：
De Wever, Olivier;Pauwels, Patrick;De Craene, Bram;Sabbah, Michele;Emami, Shahin;Redeuilh, Gerard;Gespach, Christian;Bracke, Marc;Berx, Geert
通讯作者：
Berx, Geert
DOI：
10.1073/pnas.242720499
发表时间：
2003-01-21
影响因子：
11.1
作者：
Dou, F;Netzer, WJ;Xu, HX
通讯作者：
Xu, HX