A Novel 3-Dimensional Culture System as an In Vitro Model for Studying Oral Cancer Cell Invasion
Penn collection
Degree type
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Subject
Hypoxia
Opticell chamber
Oral cancer invasion
Blotting
Western
Cell Hypoxia
Coculture Techniques
Electrophoresis
Polyacrylamide Gel
Fibroblasts
Humans
Mouth Mucosa
Mouth Neoplasms
Neoplasm Invasiveness
Plasminogen Activator Inhibitor 1
Urinary Plasminogen Activator
plasminogen activator inhibitor
urokinase receptor
article
cancer cell culture
cancer invasion
cell invasion
cell migration
controlled study
down regulation
enzyme activity
fibroblast
human
human cell
hypoxia
microscopy
mouth cancer
priority journal
protein expression
Western blotting
zymography
Dentistry
Oral Biology and Oral Pathology
Periodontics and Periodontology
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Abstract
Tissue microenvironment plays a critical role in tumour growth and invasion. This study established a novel 3-dimensional (3-D) cell invasion model for direct microscopic observation of oral cancer cell invasion into the underlying basement membrane and connective tissue stroma. A multilayer cell construct was developed using the OptiCell chamber, consisting of a lower layer of oral mucosa fibroblasts embedded in collagen gel and an overlaying upper layer of oral cancer cells. The two layers are separated by a basement membrane composed of reconstituted extracellular matrix. To verify the applicability of the cell invasion model, multilayer cell constructs of oral squamous cell carcinoma and oral mucosal fibroblasts were exposed to extrinsic urokinase-type plasminogen activator (uPA) or plasminogen activator inhibitor (PAI-1), which are known effectors of cell migration. In addition, the constructs were exposed to both normoxic and hypoxic culture conditions. Microscopic study showed that the presence of uPA enhanced cell invasion, while PAI-1 inhibited cell migration. Western blot and zymographic analysis demonstrated that hypoxia up-regulated uPA and matrix metalloproteinases (MMPs) expression and activity; conversely, PAI-1 level was down-regulated in response to hypoxic challenge as compared to normoxic condition. Our results indicated that the novel 3-D invasion model could serve as an excellent in vitro model to study cancer cell invasion and to test conditions or mediators of cellular migration. © 2005 Blackwell Publishing Ltd.