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Updated: Oct 10, 2026
A Combined 3D Tissue Engineered In Vitro/In Silico Lung Tumor Model for Predicting Drug Effectiveness in Specific Mutational Backgrounds
Published on: April 6, 2016
Reduced graphene oxide-incorporated gelatin-oxidized polymannose scaffold-based 3D Lung cancer model for mimicking
Hema Priya Manivannan1, Vishnu Priya Veeraraghavan1, K P Ameya2
1Centre of Molecular Medicine and Diagnostics (COMManD), Department of Biochemistry, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, India.
Objective:
The tumor microenvironment (TME) plays a critical role in regulating cancer progression, metastasis, and therapeutic response. Conventional two-dimensional (2D) culture systems fail to accurately recapitulate the complex native TME, highlighting the need for physiologically relevant three-dimensional (3D) in vitro models. In our previous work we have developed gelatin-oxidized polymannose-reduced graphene oxide (Gel: OPM:rGO) scaffold as a 3D lung cancer culture model. In this study, we aimed to assess its potential to support TME-associated responses compared with conventional 2D and Matrigel-based 3D culture systems.
Materials And Methods:
A549 cells cultured on the Gel: OPM:rGO scaffold were analyzed for the secretion of growth factors and inflammatory cytokines by ELISA, the expression of TME-associated and signaling pathway-related genes by qRT-PCR, and TME-associated proteins by Western blotting. Metabolic profiling was performed by measuring glucose consumption and lactate production and the results were compared with 2D and Matrigel culture systems.
Results:
Gene expression analysis revealed the upregulation of TME-associated genes, including the hypoxia marker VEGF-A, the extracellular matrix (ECM) remodeling marker MMP9, the epithelial-mesenchymal transition (EMT) marker VIM, the oncogenic marker ALK, and the growth factor receptor EGFR. Western blot analysis showed increased expression of TME-related proteins in the Gel: OPM:rGO scaffold compared with conventional 2D culture. Major signaling pathways, including mTOR, PI3K/AKT, Wnt, c-Myc, STAT3, and TNF-α, were also modulated, indicating physiologically relevant cellular responses. Metabolic profiling in Gel: OPM:rGO culture supernatant demonstrated enhanced glucose consumption and lactate production, consistent with the Warburg effect and tumor-associated metabolic reprogramming. Overall, gene and protein expression profiles in the Gel: OPM:rGO scaffold exhibited trends similar to those observed in Matrigel-based 3D culture.
Conclusion:
Collectively, these findings demonstrate that the Gel: OPM:rGO scaffold is a robust and physiologically relevant 3D lung cancer model. As a potential 3D culture platform that complements Matrigel-based models, it provides a valuable in vitro platform for investigating tumor-associated cellular and molecular responses, cancer progression, and preclinical anticancer drug evaluation.
