Abstract
Abstract The hollow fibre assay (HFA) facilitates the short-term assessment of cancer drug efficacy in vivo, bridging in vitro screening methods and more complex xenograft in vivo models. However a drawback of this model is the need for end-stage cell recovery and assessment in order to determine the test compound effects. An additional limitation of this system is the use of a homogenous population of epithelial cells, which is not representative of the solid tumor micro-environment, a critical component in tumor progression and the development of drug resistance. The aims of our current research were to recapitulate the human tumor micro-environment within the hollow fibre model and apply a real-time bio-imaging component to enable the non-invasive analysis of drug responsiveness and toxicity in addition to evaluate markers of tumor progression, which was confirmed histologically. Using epithelial tumor cells stably expressing fluorescent/bioluminescent reporter genes, we have investigated cell viability and the response to cytotoxic treatment in the presence and absence of human mesenchymal stem cells (MSCs) and cancer-associated fibroblasts (CAFs) both within in vitro cell cultures and the HFA. Real-time imaging of bioluminescent tumor cells demonstrated that these cells can actively proliferate for up to 16 days within the hollow fibres. In the presence of MSCs or CAFs tumor growth was significantly enhanced compared with tumor cells alone and immuno—staining analysis revealed stromal deposits around the periphery of the fibre with MSCs acquiring a ‘myofibroblastic’ phenotype with prolonged culture, resulting in growth promoting effects similar to CAFs. Tumor cells expressing dual reporters with luciferase under the control of 3 hypoxia response elements (HREs) and RFP driven by a CMV promoter confirmed the development of hypoxia within the HFA and enabled further investigation of cancer stem cell development and epithelial:mesenchymal transition. This re-capitulation of the human tumor micro-environment provides a relevant model by which to identify and characterise novel anti-cancer compounds. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C247.
Published Version
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