Focal adhesion kinase is activated in invading fibrosarcoma cells and regulates metastasis
Focal adhesion kinase is activated in invading fibrosarcoma cells and regulates metastasis
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DOI：
10.1007/s10585-005-3733-6
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发表时间：
2005-11-01
影响因子：
4
通讯作者：
Iwamoto, Y
中科院分区：
文献类型：
--
作者：
Hanada, M;Tanaka, K;Iwamoto, Y
粘着斑激酶（FAK）是一种非受体酪氨酸激酶，在几种人类癌症中过表达，并诱导培养细胞的存活、增殖和运动。FAK的磷酸化在体外已被广泛研究，但其在体内肿瘤侵袭过程中的调节知之甚少。在本研究中，绿色荧光蛋白（GFP）在侵袭性小鼠纤维肉瘤细胞系中稳定表达，用于区分肿瘤和正常细胞。在荧光显微镜下，肿瘤呈高荧光，肿瘤和正常组织之间的边界清晰。使用这种侵袭模型，我们显示了pY 397-FAK表达在肿瘤浸润边缘的定位。我们在体内用三维胶原凝胶中的肿瘤组织培养方法再现了局部侵袭。在体外条件下，在侵袭性纤维肉瘤细胞中FAK的磷酸化也上调。在2472细胞中FAK C-末端结构域称为FRNK（FAK相关非激酶）的表达降低FAK磷酸化而不改变总FAK水平。FRNK可抑制2472细胞的运动能力，降低其体外侵袭能力。虽然FRNK不影响细胞生长，但它抑制了同系小鼠的实验转移。这些结果表明，FAK的磷酸化可能在侵袭纤维肉瘤细胞中特异性上调，并调节其侵袭和转移。
Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase that is overexpressed in several human cancers, and induces survival, proliferation and motility of cells in culture. Phosphorylation of FAK has been studied extensively in vitro, but little is known about its regulation during tumor invasion in vivo. In the current study, green fluorescent protein (GFP) was expressed stably in an invasive murine fibrosarcoma cell line for the purpose of discrimination between tumor and normal cells. Under fluorescence microscopy, the tumor was highly fluorescent, and the margin between the tumor and normal tissue was clearly demarcated. Using this invasion model, we showed localization of pY397-FAK expression in the infiltrative edge of tumors. We reproduced local invasion in vivo using a tumor tissue culture method in a three dimensional collagen gel. Phosphorylation of FAK is also upregulated in invading fibrosarcoma cells under in vitro conditions. Expression of the FAK C-terminal domain termed FRNK (FAK-related non-kinase) in 2472 cells decreased FAK phosphorylation without changing total FAK levels. FRNK inhibited the motility of 2472 cells, and reduced invasion in vitro. Although FRNK did not affect cell growth, it inhibited experimental metastases in syngenic mice. These results demonstrate that the phosphorylation of FAK might be specifically upregulated in invading fibrosarcoma cells and regulate their invasion and metastasis.