Real-time Study of E-Cadherin and Membrane Dynamics in Living Animals: Implications for Disease Modeling and Drug Development
Real-time Study of E-Cadherin and Membrane Dynamics in Living Animals: Implications for Disease Modeling and Drug Development
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DOI：
10.1158/0008-5472.can-08-4308
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发表时间：
2009-04-01
期刊：
影响因子：
11.2
通讯作者：
Anderson, Kurt I.
中科院分区：
文献类型：
--
作者：
Serrels, Alan;Timpson, Paul;Anderson, Kurt I.
肿瘤细胞侵袭和转移的能力需要与邻近细胞和细胞外基质的相互作用的失调。癌症生物学的一个主要挑战是观察参与这一过程的蛋白质在其功能和生理背景下的动力学。在这里，我们第一次使用光漂白和光活化来比较细胞粘附和质膜探针在体外和在小鼠（体内）生长的肿瘤中的移动性。我们发现两个关键分子，肿瘤抑制因子E-钙粘蛋白和H-Ras的膜靶向序列在体外和体内恢复动力学之间的差异。我们的数据表明，E-钙粘蛋白的动力学是显着更快，在体内培养的细胞相比，E-钙粘蛋白的比例稳定在细胞-细胞连接是显着更高的体内，和E-钙粘蛋白的流动性与细胞迁移。此外，定量成像使我们能够评估使用达沙替尼（一种临床批准的Src抑制剂）的治疗干预对E-钙粘蛋白动力学的影响，并显示出体内药物治疗疗效的明显差异。我们的研究结果首次显示了光漂白和光活化在活体动物动态生物标志物分析中的实用性。此外，这项工作突出了体外和体内分子动力学的关键差异，这对使用培养的疾病模型作为活组织的替代物具有重要意义。[Cancer Res 2009;69（7）：2714-9]
The ability of tumor cells to invade and metastasize requires deregulation of interactions with adjacent cells and the extracellular matrix. A major challenge of cancer biology is to observe the dynamics of the proteins involved in this process in their functional and physiologic context. Here, for the first time, we have used photobleaching and photoactivation to compare the mobility of cell adhesion and plasma membrane probes in vitro and in tumors grown in mice (in vivo). We find differences between in vitro and in vivo recovery dynamics of two key molecules, the tumor suppressor E-cadherin and the membrane-targeting sequence of H-Ras. Our data show that E-cadherin dynamics are significantly faster in vivo compared with cultured cells, that the ratio of E-cadherin stabilized in cell-cell junctions is significantly higher in vivo, and that E-cadherin mobility correlates with cell migration. Moreover, quantitative imaging has allowed us to assess the effects of therapeutic intervention on E-cadherin dynamics using dasatinib, a clinically approved Src inhibitor, and show clear differences in the efficacy of drug treatment in vivo. Our results show for the first time the utility of photobleaching and photoactivation in the analysis of dynamic biomarkers in living animals. Furthermore, this work highlights critical differences in molecular dynamics in vitro and in vivo, which have important implications for the use of cultured disease models as surrogates for living tissue. [Cancer Res 2009;69(7):2714-9]