Culturing substrates influence the morphological, mechanical and biochemical features of lung adenocarcinoma cells cultured in 2D or 3D. (February 2018)

• Record Type: Journal Article
• Authors: Prina-Mello, Adriele; Jain, Namrata; Liu, Baiyun; Kilpatrick, Jason I.; Tutty, Melissa A.; Bell, Alan P.; Jarvis, Suzanne P; Volkov, Yuri; Movia, Dania
• Is Part Of: Tissue & cell. Volume 50(2018)
• Page Start: 15
• Page End: 30
• Publication Date: 2018-02
• Subjects: 3D cell cultures -- Non-small-cell lung cancer -- Mechanical properties -- Hydrogel-based cell cultures -- Matrigel -- PuraMatrix™; Cytology -- Periodicals; 571.5
• DOI: 10.1016/j.tice.2017.11.003
• Languages: English
• ISSNs: 0040-8166

Abstract

Graphical abstract: Highlights: Three-dimensional spatial organization influences A549 cells' shape. Substrate stiffness influences cytoskeleton organization and cell roughness. In 3D, cell roughness is associated with invasion, but not to EMT biomarkers. In 3D, IL-8 promotes tumour progression through PI3K/Akt and MAPK/Erk1/2 pathways. Abstract: 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.