Arora Leena, Kalia Moyna, Dasgupta Suman, Singh Navneet, Verma Anita K, Pal Durba
Department of Biomedical Engineering, Indian Institute of Technology Ropar, Punjab, India.
Department of Molecular Biology & Biotechnology, Tezpur University, Assam, India.
Front Oncol. 2022 Jun 28;12:881207. doi: 10.3389/fonc.2022.881207. eCollection 2022.
Heterogeneity is a characteristic feature of solid tumors. Intra-tumor heterogeneity includes phenotypic diversity, epigenetic abnormalities, cell proliferation, and plasticity that eventually drives disease progression. Studying tumor heterogeneity in 2D culture is challenging as it cannot simulate the microenvironmental features, such as hypoxia, nutrient unavailability, and cell-ECM interactions. We propose the development of multicellular (tri-culture) 3D spheroids using a hanging drop method to study the non-tumorigenic (BEAS-2B) vs. tumorigenic NSCLC (A549/NCI-H460)cells' interaction with lung fibroblasts (MRC-5) and monocytes (THP-1). Unlike the non-tumorigenic model, the tumorigenic 3D spheroids show significant induction of cell proliferation, hypoxia, pluripotency markers, notable activation of cancer-associated fibroblasts, and tumor-associated macrophages. CD68+ macrophages isolated from tumorigenic spheroids exhibited profound induction of phenotypic endothelial characteristics. The results are zebrafish tumor xenograft model and by using human patient samples. This multicellular 3D tumor model is a promising tool to study tumor-stroma interaction and cellular plasticity, targeting tumor heterogeneity, and facilitating cancer therapy success against NSCLC.
异质性是实体瘤的一个特征。肿瘤内异质性包括表型多样性、表观遗传异常、细胞增殖和可塑性，这些最终推动疾病进展。在二维培养中研究肿瘤异质性具有挑战性，因为它无法模拟微环境特征，如缺氧、营养物质缺乏以及细胞与细胞外基质的相互作用。我们建议使用悬滴法开发多细胞（三培养）三维球体，以研究非致瘤性（BEAS-2B）与致瘤性非小细胞肺癌（A549/NCI-H460）细胞与肺成纤维细胞（MRC-5）和单核细胞（THP-1）的相互作用。与非致瘤性模型不同，致瘤性三维球体显示出细胞增殖、缺氧、多能性标志物的显著诱导，癌症相关成纤维细胞和肿瘤相关巨噬细胞的明显激活。从致瘤性球体中分离出的CD68+巨噬细胞表现出表型内皮特征的深度诱导。这些结果通过斑马鱼肿瘤异种移植模型并使用人类患者样本得到验证。这种多细胞三维肿瘤模型是研究肿瘤-基质相互作用和细胞可塑性、靶向肿瘤异质性以及促进非小细胞肺癌癌症治疗成功的一种有前景的工具。
