Department of Cell Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-9039, United States.
Department of Cell Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-9039, United States.
Acta Biomater. 2019 Dec;100:213-222. doi: 10.1016/j.actbio.2019.09.033. Epub 2019 Sep 25.
Current 3D culture models to study colorectal cancer lack architectural support and signaling proteins provided by the tissue extracellular matrix (ECM) which may influence cell behavior and cancer progression. Therefore, the ability to study cancer cells in the context of a matrix that is physiologically more relevant and to understand how the ECM affects cancer progression has been understudied. To address this, we developed an ex-vivo 3D system, provided by intact wild type (WT) and colon cancer susceptible decellularized mouse colons (DMC), to support the growth of human cancer cells. DMC are free of viable cells but still contain extracellular matrix proteins including subsets of collagens. Stiffness, an important mechanical property, is also maintained in DMCs. Importantly, we observed that the DMC is permissive for cell proliferation and differentiation of a human colon cancer cell line (HT-29). Notably, the ability of cells in the WT DMC to differentiate was also greater when compared to Matrigel™, an extracellular matrix extract from a mouse tumor cell line. Additionally, we observed in invasion assays that DMC obtained from polyps from a colon cancer susceptible mouse model facilitated increased cell migration/invasion of colorectal cancer cells and immortalized non-tumor colonic epithelial cells compared to DMC from WT mice. Finally, using mass spectrometry, we identified extracellular matrix proteins that are more abundant in DMC from a colorectal cancer mouse model compared to age and sex-matched WT mice. We propose that these abundantly expressed proteins in the tumor microenvironment are potentially involved in colorectal cancer progression. STATEMENT OF SIGNIFICANCE: Decellularized matrices, when properly produced, are attractive biomaterials for tissue regeneration and replacement. We show here that the mouse decellularized matrices can also be repurposed to elucidate how the extracellular matrix influences human cell behavior and cancer progression. To do this we produce decellularized matrices, from mice colonic tissue, that have preserved tissue mechanical and structural properties. We demonstrate that the matrix better supports the differentiation of HT-29 cells, a colonic cancer cell line, compared to Matrigel™. Additionally, we show that the extracellular matrix contributes to colon cancer progression via invasion assays using extracellular matrix extracts. Finally, we use mass spectrometry to identify ECM proteins that are more abundant in colonic polyps compared to adjacent tissue regions. This model system may have therapeutic implications for colorectal cancer patients.
目前用于研究结直肠癌的 3D 培养模型缺乏组织细胞外基质（ECM）提供的结构支撑和信号蛋白，而这些蛋白可能会影响细胞行为和癌症进展。因此，研究在更接近生理状态的基质中癌症细胞的能力，以及了解 ECM 如何影响癌症进展的研究还很不足。为了解决这个问题，我们开发了一种离体 3D 系统，该系统由完整的野生型（WT）和易发生结肠癌的去细胞化小鼠结肠（DMC）提供，以支持人类癌细胞的生长。DMC 不含活细胞，但仍含有细胞外基质蛋白，包括部分胶原蛋白。DMC 还保留了硬度这一重要的力学特性。重要的是，我们观察到 DMC 允许人结肠癌细胞系（HT-29）增殖和分化。值得注意的是，与从一种小鼠肿瘤细胞系提取的细胞外基质提取物 MatrigelTM 相比，WT-DMC 中的细胞分化能力也更强。此外，我们在侵袭实验中观察到，与 WT 小鼠来源的 DMC 相比，来自易发生结肠癌的小鼠模型的息肉来源的 DMC 促进了结直肠癌细胞和永生化非肿瘤结肠上皮细胞的迁移/侵袭。最后，通过质谱分析，我们鉴定出与年龄和性别匹配的 WT 小鼠相比，在结直肠癌小鼠模型来源的 DMC 中更丰富的细胞外基质蛋白。我们提出，这些在肿瘤微环境中丰富表达的蛋白质可能与结直肠癌的进展有关。
去细胞化基质在组织再生和替代方面具有吸引力，是一种很有前途的生物材料。我们在这里表明，当正确制备时，小鼠去细胞化基质也可以重新用于阐明细胞外基质如何影响人类细胞行为和癌症进展。为此，我们从小鼠结肠组织中制备保留了组织力学和结构特性的去细胞化基质。我们证明该基质比 MatrigelTM 更能支持 HT-29 细胞（一种结肠癌细胞系）的分化。此外，我们通过使用细胞外基质提取物进行侵袭实验表明，细胞外基质有助于结肠癌的进展。最后，我们使用质谱分析鉴定出在结直肠息肉中比相邻组织区域更丰富的 ECM 蛋白。该模型系统可能对结直肠癌患者具有治疗意义。
