Nawrocki Raby B, Polette M, Gilles C, Clavel C, Strumane K, Matos M, Zahm J M, Van Roy F, Bonnet N, Birembaut P
INSERM U514, CHU Maison Blanche, Reims, France.
Int J Cancer. 2001 Sep 1;93(5):644-52. doi: 10.1002/ijc.1380.
Tumor progression requires the dispersion of epithelial cells from neoplastic clusters and cell invasion of adjacent stromal connective tissue. Aiming at demonstrating the precise relationships between cell dispersion and cell invasion, related respectively to expression of E-cadherin/catenin complex and matrix metalloproteinases (MMPs), we developed an original in vitro model of cell dispersion analysis. Our study reports the validation of this model that allowed us to analyze and quantify the cell cohesion level by means of time-lapse videomicroscopy and computer analysis based on the observation of spatial and temporal cell distribution. Our model was able to distinguish 2 groups among different human bronchial and mammary epithelial cells previously characterized for the expression of E-cadherin/catenin complex and MMPs and their invasive capacity in the Boyden chamber assay. The first group (16HBE14o(-), MCF-7, T47D) that expressed membranous E-cadherin and beta-catenin, and was negative for MMP-2 expression and non-invasive, displayed a highly cohesive pattern corresponding to a cluster spatial distribution. The second group (Beas2B, BZR, BZR-T33, MDA-MB-231, MDA-MB-435, BT549 and HS578T) that was invasive and showed lack of expression of E-cadherin and a cytoplasmic redistribution of beta-catenin, displayed a dispersed pattern corresponding to a random spatial distribution. Downregulation of E-cadherin by a blocking antibody induced a more random distribution. Conversely, expression of E-cadherin by cDNA transfection induced a cluster distribution. Moreover, tumor cell lines that co-expressed MT1-MMP and MMP-2 (Beas2B, BZR, BZR-T33, MDA-MB-435, BT549 and HS578T) showed a more dispersed pattern than tumor cell lines that did not express MMP-2 (MDA-MB-231). In conclusion, we demonstrated that the spatial group behavior of cell lines, i.e., their cohesion/dispersion ability, reflects their invasive properties. Thus, this model of cell dispersion analysis may represent a new test to measure tumor cell aggressiveness.
肿瘤进展需要上皮细胞从肿瘤细胞簇中分散出来，并侵入相邻的基质结缔组织。为了阐明细胞分散与细胞侵袭之间分别与E-钙黏蛋白/连环蛋白复合体和基质金属蛋白酶（MMPs）表达相关的精确关系，我们开发了一种原创的细胞分散分析体外模型。我们的研究报告了该模型的验证情况，该模型使我们能够通过延时视频显微镜以及基于对细胞时空分布观察的计算机分析来分析和量化细胞黏附水平。我们的模型能够在先前已根据E-钙黏蛋白/连环蛋白复合体和MMPs的表达及其在博伊登小室试验中的侵袭能力进行表征的不同人支气管和乳腺上皮细胞中区分出两组。第一组（16HBE14o(-)、MCF-7、T47D）表达膜性E-钙黏蛋白和β-连环蛋白，MMP-2表达呈阴性且无侵袭性，呈现出与细胞簇空间分布相对应的高度黏附模式。第二组（Beas2B、BZR、BZR-T33、MDA-MB-231、MDA-MB-435、BT549和HS578T）具有侵袭性，E-钙黏蛋白表达缺失且β-连环蛋白发生细胞质重新分布，呈现出与随机空间分布相对应的分散模式。用阻断抗体下调E-钙黏蛋白会诱导出更随机的分布。相反，通过cDNA转染表达E-钙黏蛋白会诱导出细胞簇分布。此外，共表达MT1-MMP和MMP-2的肿瘤细胞系（Beas2B、BZR、BZR-T33、MDA-MB-435、BT549和HS578T）比不表达MMP-2的肿瘤细胞系（MDA-MB-231）呈现出更分散的模式。总之，我们证明了细胞系的空间群体行为，即它们的黏附/分散能力，反映了它们的侵袭特性。因此，这种细胞分散分析模型可能代表了一种测量肿瘤细胞侵袭性的新测试方法。
