Shenzhen Key Laboratory of Gastrointestinal Cancer Translational Research, Department of Oncology, Peking University Shenzhen Hospital, Cancer Institute of Shenzhen-PKU-HKUST Medical Center, Shenzhen, P.R. China.
Department of Radiation Oncology and Peking University Shenzhen Hospital, Shenzhen, P.R. China.
DNA Cell Biol. 2021 Jul;40(7):869-880. doi: 10.1089/dna.2020.6467. Epub 2021 Jun 28.
Non-small cell lung cancer is the most common type of lung cancer and is a frequent cause of death. In our research, A549 and SK-MES-1 were used to assess the effect of three-dimensional (3D) culture compared to that of two-dimensional (2D) monolayers in cell proliferation, migration, and invasion, response to chemotherapy, as well as the expression of epithelial to mesenchymal transition (EMT) and cancer stem cell (CSC)-related markers. As tadalafil is a phosphodiesterase type 5 (PDE5) inhibitor with the potential to target CSC maintenance in multiple cancer cell lines, we assessed its function in 3D culture and detected the downstream pathway genes. We compared the viability and proliferation capacity of A549 and SK-MES-1 cells in 2D and 3D culture by cell counting kit (CCK)-8, foci formation, and Live/Dead double stain (Operetta CLS High content screening). Migration, invasion, and other functions were also assessed. To elucidate the underlying mechanisms, the expression of EMT and CSC markers was analyzed by quantitative real-time PCR (qPCR) and Western blot. A549 and SK-MES-1 cells formed spheroids heterogeneous in shape and size. In our 3D spheroid systems, cells went through EMT, and were also capacitated with higher stemness and chemoresistance. Combination use of tadalafil and cisplatin showed higher chemotherapy efficiency in the 3D model, compared to that of the 2D cell culture. Our research aims at the notable differences between these two cellular systems in terms of cell functions, EMT, and stemness-associated gene expression and chemo-response. We showed that a commonly used drug, tadalafil, showed more pronounced inhibitory effects when cells were cultured in the 3D model. Since the 3D culture system could imitate the behavior of cancer cells within the tumor, we advocate that this system is superior to traditional 2D culture and should be used in the investigation of new therapeutic compounds.
非小细胞肺癌是最常见的肺癌类型，也是导致死亡的常见原因。在我们的研究中，使用 A549 和 SK-MES-1 细胞来评估三维（3D）培养与二维（2D）单层培养相比，在细胞增殖、迁移和侵袭、对化疗的反应以及上皮间质转化（EMT）和癌症干细胞（CSC）相关标志物的表达方面的差异。由于他达拉非是一种磷酸二酯酶 5（PDE5）抑制剂，具有靶向多种癌细胞系中 CSC 维持的潜力，我们评估了它在 3D 培养中的功能，并检测了下游通路基因。我们通过细胞计数试剂盒（CCK-8）、焦点形成和 Live/Dead 双重染色（Operetta CLS 高内涵筛选）比较了 A549 和 SK-MES-1 细胞在 2D 和 3D 培养中的活力和增殖能力。还评估了迁移、侵袭和其他功能。为了阐明潜在的机制，通过实时定量 PCR（qPCR）和 Western blot 分析了 EMT 和 CSC 标志物的表达。A549 和 SK-MES-1 细胞形成了形状和大小不均匀的球体。在我们的 3D 球体系统中，细胞经历了 EMT，并具有更高的干性和化疗耐药性。与 2D 细胞培养相比，他达拉非和顺铂的联合使用在 3D 模型中显示出更高的化疗效率。我们的研究旨在研究这两种细胞系统在细胞功能、EMT 和干性相关基因表达和化疗反应方面的显著差异。我们表明，一种常用药物他达拉非在细胞在 3D 模型中培养时显示出更明显的抑制作用。由于 3D 培养系统可以模拟肿瘤内癌细胞的行为，我们主张该系统优于传统的 2D 培养，应该用于新治疗化合物的研究。
