The microenvironment determines the breast cancer cells' phenotype: organization of MCF7 cells in 3D cultures.
The microenvironment determines the breast cancer cells' phenotype: organization of MCF7 cells in 3D cultures.
复制标题
DOI：
10.1186/1471-2407-10-263
复制
发表时间：
2010-06-07
期刊：
影响因子：
3.8
通讯作者：
Sonnenschein C
中科院分区：
文献类型：
--
作者：
Krause S;Maffini MV;Soto AM;Sonnenschein C
基质-上皮相互作用介导乳腺发育以及乳腺癌的发生和发展。在本研究中，我们开发了三维（3D）体外模型来研究乳腺癌组织的组织结构和微环境在表型确定中的作用。人乳腺癌MCF 7细胞单独生长或与原代人乳腺成纤维细胞共培养。将细胞包埋在含有I型胶原或重组基底膜蛋白和I型胶原的组合的基质中。培养进行长达6周。对于每个时间点（1-6周），将凝胶固定并处理用于组织学，并整体封固用于共聚焦显微镜评价。上皮结构的特点，利用免疫组织化学技术，其面积和增殖指数进行了测量，使用计算机形态学分析。通过ANOVA、Dunnett T3事后检验和卡方分析组间的统计学差异。大多数在胶原基质中单独生长的MCF 7细胞在前两周内死亡;那些存活下来的细胞组织成大的，圆形的和固体的簇。胶原蛋白凝胶中成纤维细胞的存在减少了MCF 7细胞死亡，诱导了细胞极性，并形成了圆形和细长的上皮结构，含有管腔。向胶原凝胶中添加重构基底膜本身也对MCF 7细胞具有存活和组织影响。无论是否存在成纤维细胞，MCF 7细胞均极化并形成管腔。将成纤维细胞添加到含有重构基底膜和胶原蛋白的凝胶中诱导了细长结构的形成。我们的结果表明，含有I型胶原和重建基底膜的基质以及正常乳腺成纤维细胞的存在构成了诱导几乎完全的肿瘤表型逆转的最小允许微环境。这些人类乳腺3D组织形态发生模型有望成为研究组织相互作用、治疗筛选和药物靶点验证的可靠工具。
Stromal-epithelial interactions mediate breast development, and the initiation and progression of breast cancer. In the present study, we developed 3-dimensional (3D) in vitro models to study breast cancer tissue organization and the role of the microenvironment in phenotypic determination. The human breast cancer MCF7 cells were grown alone or co-cultured with primary human breast fibroblasts. Cells were embedded in matrices containing either type I collagen or a combination of reconstituted basement membrane proteins and type I collagen. The cultures were carried out for up to 6 weeks. For every time point (1-6 weeks), the gels were fixed and processed for histology, and whole-mounted for confocal microscopy evaluation. The epithelial structures were characterized utilizing immunohistochemical techniques; their area and proliferation index were measured using computerized morphometric analysis. Statistical differences between groups were analyzed by ANOVA, Dunnett's T3 post-hoc test and chi-square. Most of the MCF7 cells grown alone within a collagen matrix died during the first two weeks; those that survived organized into large, round and solid clusters. The presence of fibroblasts in collagen gels reduced MCF7 cell death, induced cell polarity, and the formation of round and elongated epithelial structures containing a lumen. The addition of reconstituted basement membrane to collagen gels by itself had also survival and organizational effects on the MCF7 cells. Regardless of the presence of fibroblasts, the MCF7 cells both polarized and formed a lumen. The addition of fibroblasts to the gel containing reconstituted basement membrane and collagen induced the formation of elongated structures. Our results indicate that a matrix containing both type I collagen and reconstituted basement membrane, and the presence of normal breast fibroblasts constitute the minimal permissive microenvironment to induce near-complete tumor phenotype reversion. These human breast 3D tissue morphogenesis models promise to become reliable tools for studying tissue interactions, therapeutic screening and drug target validation.
登录
查看更多内容
影响因子：
2.5
作者：
Howard, BA;Gusterson, BA
通讯作者：
Gusterson, BA
影响因子：
48
作者：
Fischbach, Claudia;Chen, Ruth;Mooney, David J.
通讯作者：
Mooney, David J.
影响因子：
2.5
作者：
Kasemeier-Kulesa, Jennifer C.;Teddy, Jessica M.;Postovit, Lynne-Marie;Seftor, Elisabeth A.;Seftor, Richard E. B.;Hendrix, Mary J. C.;Kulesa, Paul M.
通讯作者：
Kulesa, Paul M.
影响因子：
3
作者：
Krause, Silva;Maffini, Maricel V.;Sonnenschein, Carlos
通讯作者：
Sonnenschein, Carlos
影响因子：
64.8
作者：
Liotta, LA;Kohn, EC
通讯作者：
Kohn, EC