title: A microfluidic coculture and multiphoton FAD analysis assay provides insight into the influence of the bone microenvironment on prostate cancer cells.
abstract: In prostate cancer, bone is a frequent site of metastasis; however, the molecular mechanisms of this tumor tropism remain unclear. Here, we integrate a microfluidic coculture platform with multi-photon imaging based techniques to assess both phenotypic cell behavior and FAD fluorescence intensity and fluorescence lifetime in the same cell. This platform combines two independent assays normally performed with two different cell populations into a single device, allowing us to simultaneously assess both phenotypic cell behavior and enzyme activity. We observed that the osteotropic prostate cancer cell line (C4-2B), when in a coculture with bone marrow stromal cells (MC3T3-E1), has increased protrusive phenotype and increased total and protein-bound FAD compared to its parent cell line (LNCaP). We hypothesized that an increase in ROS-generating APAO activity may be responsible for these effects, and found that the effects were decreased in the presence of the antioxidant N-Acetyl Cysteine (NAC). This suggests that an ROS-related signaling mechanism at the bone metastatic site may be correlated with and play a role in increased invasion of metastasizing prostate cancer cells. The studies performed using this combined platform will lead to new insights into the mechanisms that drive prostate cancer metastasis.
authors.affiliation: Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin, Madison, WI, USA.
authors.forename: Lauren L
authors.initials: LL
authors.lastname: Bischel
authors.affiliation: Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin, Madison, WI, USA.
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Casavant
authors.affiliation: Laboratory for Optical and Computational Instrumentation, University of Wisconsin, Madison, WI, USA.
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Young
authors.affiliation: Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin, Madison, WI, USA.|Laboratory for Optical and Computational Instrumentation, University of Wisconsin, Madison, WI, USA.
authors.forename: Kevin W
authors.initials: KW
authors.lastname: Eliceiri
authors.affiliation: Laboratory for Optical and Computational Instrumentation, University of Wisconsin, Madison, WI, USA.|Department of Medicine, University of Wisconsin Carbone Cancer Center, Madison, WI, USA.
authors.forename: Hirak S
authors.initials: HS
authors.lastname: Basu
authors.affiliation: Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin, Madison, WI, USA.|Laboratory for Optical and Computational Instrumentation, University of Wisconsin, Madison, WI, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism*; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D044085:Microfluidics; D036641:Microscopy, Fluorescence, Multiphoton; D011471:Prostatic Neoplasms / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D017382:Reactive Oxygen Species; D005182:Flavin-Adenine Dinucleotide; D000111:Acetylcysteine