A synthetic matrix with independently tunable biochemistry and mechanical properties to study epithelial morphogenesis and EMT in a lung adenocarcinoma model.
A synthetic matrix with independently tunable biochemistry and mechanical properties to study epithelial morphogenesis and EMT in a lung adenocarcinoma model.
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DOI：
10.1158/0008-5472.can-12-0895
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发表时间：
2012-11-15
期刊：
影响因子：
11.2
通讯作者：
West JL
中科院分区：
文献类型：
--
作者：
Gill BJ;Gibbons DL;Roudsari LC;Saik JE;Rizvi ZH;Roybal JD;Kurie JM;West JL
更好地了解影响转移的肿瘤细胞外基质环境的生物物理和生化线索可能对新的癌症治疗具有重要意义。对这个问题的初步探索使用了自然衍生的蛋白质基质，这些基质具有可变性，对基质生物化学的控制较差，并且无法修改基质的生物化学和力学。在这里，我们报道了一种主要由聚乙二醇（PEG）或生物活性肽修饰的合成聚合物基支架的使用，用于研究小鼠肺腺癌模型。在这项研究中，我们重点研究了基质来源对转移细胞系（344SQ）上皮形态发生的影响，该细胞系含有Kras和p53（trp53）突变，容易发生依赖microRNA-200 （miR-200）的上皮-间质转化（EMT）和转移。改性的PEG水凝胶具有生物特异性细胞粘附和细胞介导的蛋白水解降解，具有可独立调节的基质刚度。344SQ被包裹在生物活性肽修饰的基质金属蛋白酶可降解的PEG水凝胶中，形成了流明的上皮球，与在Matrigel中三维培养所见的相似。基质硬度和细胞黏附配体浓度的改变会显著影响上皮的形态发生，表现在流明化程度、球内细胞凋亡和增殖模式以及上皮极性标记物的表达上。无论基质成分如何，暴露于TGF-β都会导致上皮形态特征的丧失，EMT标记基因的表达发生变化，mir-200水平下降与EMT一致。我们的发现有助于阐明影响上皮形态发生的基质来源线索，并强调这种合成基质模拟工具在癌症生物学中的潜在效用。
Better understanding of the biophysical and biochemical cues of the tumor extracellular matrix environment that influence metastasis may have important implications for new cancer therapeutics. Initial exploration into this question has used naturally derived protein matrices that suffer from variability, poor control over matrix biochemistry, and inability to modify the matrix biochemistry and mechanics. Here, we report the use of a synthetic polymer-based scaffold composed primarily of poly(ethylene glycol), or PEG, modified with bioactive peptides to study murine models of lung adenocarcinoma. In this study, we focus on matrix-derived influences on epithelial morphogenesis of a metastatic cell line (344SQ) that harbors mutations in Kras and p53(trp53) and is prone to a microRNA-200 (miR-200)–dependent epithelial–mesenchymal transition (EMT) and metastasis. The modified PEG hydrogels feature biospecific cell adhesion and cell-mediated proteolytic degradation with independently adjustable matrix stiffness. 344SQ encapsulated in bioactive peptide-modified, matrix metalloproteinase–degradable PEG hydrogels formed lumenized epithelial spheres comparable to that seen with three-dimensional culture in Matrigel. Altering both matrix stiffness and the concentration of cell-adhesive ligand significantly influenced epithelial morphogenesis as manifest by differences in the extent of lumenization, in patterns of intrasphere apoptosis and proliferation, and in expression of epithelial polarity markers. Regardless of matrix composition, exposure to TGF-β induced a loss of epithelial morphologic features, shift in expression of EMT marker genes, and decrease in mir-200 levels consistent with EMT. Our findings help illuminate matrix-derived cues that influence epithelial morphogenesis and highlight the potential utility that this synthetic matrix-mimetic tool has for cancer biology.