Prina-Mello Adriele, Jain Namrata, Liu Baiyun, Kilpatrick Jason I, Tutty Melissa A, Bell Alan P, Jarvis Suzanne P, Volkov Yuri, Movia Dania
CRANN Institute and AMBER Centre, Trinity College Dublin, Ireland; Laboratory for Biological Characterization of Advanced Materials (LBCAM), Trinity Translational Medicine Institute (TTMI), Trinity College Dublin, Ireland; Department of Clinical Medicine, School of Medicine, Trinity College Dublin, Ireland.
CRANN Institute and AMBER Centre, Trinity College Dublin, Ireland.
Tissue Cell. 2018 Feb;50:15-30. doi: 10.1016/j.tice.2017.11.003. Epub 2017 Dec 1.
Alternative models such as three-dimensional (3D) cell cultures represent a distinct milestone towards capturing the realities of cancer biology in vitro and reduce animal experimentation in the preclinical stage of drug discovery. Significant work remains to be done to understand how substrates used in in vitro alternatives influence cancer cells phenotype and drug efficacy responses, so that to accurately link such models to specific in vivo disease scenarios. Our study describes how the morphological, mechanical and biochemical properties of adenocarcinoma (A549) cells change in response to a 3D environment and varying substrates. Confocal Laser Scanning (LSCM), He-Ion (HIM) and Atomic Force (AFM) microscopies, supported by ELISA and Western blotting, were used. These techniques enabled us to evaluate the shape, cytoskeletal organization, roughness, stiffness and biochemical signatures of cells grown within soft 3D matrices (PuraMatrix™ and Matrigel™), and to compare them to those of cells cultured on two-dimensional glass substrates. Cell cultures are also characterized for their biological response to docetaxel, a taxane-type drug used in Non-Small-Cell Lung Cancer (NSCLC) treatment. Our results offer an advanced biophysical insight into the properties and potential application of 3D cultures of A549 cells as in vitro alternatives in lung cancer research.
诸如三维（3D）细胞培养等替代模型是在体外捕捉癌症生物学真实情况并减少药物发现临床前阶段动物实验方面的一个重要里程碑。要了解体外替代模型中使用的基质如何影响癌细胞表型和药物疗效反应，仍有大量工作要做，以便将此类模型与特定的体内疾病情况准确联系起来。我们的研究描述了腺癌（A549）细胞的形态、机械和生化特性如何响应3D环境和不同基质而发生变化。使用了共聚焦激光扫描（LSCM）、氦离子（HIM）和原子力（AFM）显微镜，并辅以酶联免疫吸附测定（ELISA）和蛋白质印迹法。这些技术使我们能够评估在柔软的3D基质（PuraMatrix™和基质胶™）中生长的细胞的形状、细胞骨架组织、粗糙度、硬度和生化特征，并将它们与在二维玻璃基质上培养的细胞进行比较。细胞培养物还针对其对多西他赛（一种用于非小细胞肺癌（NSCLC）治疗的紫杉烷类药物）的生物学反应进行了表征。我们的结果为A549细胞3D培养物在肺癌研究中作为体外替代模型的特性和潜在应用提供了深入的生物物理见解。
