A microfluidic coculture and multiphoton FAD analysis assay provides insight into the influence of the bone microenvironment on prostate cancer cells
A microfluidic coculture and multiphoton FAD analysis assay provides insight into the influence of the bone microenvironment on prostate cancer cells
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DOI：
10.1039/c3ib40240a
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发表时间：
2014-01-01
影响因子：
2.5
通讯作者：
Beebe, David J.
中科院分区：
文献类型：
--
作者：
Bischel, Lauren L.;Casavant, Benjamin P.;Beebe, David J.
在前列腺癌中，骨是常见的转移部位。然而，这种肿瘤趋向性的分子机制仍不清楚。在这里，我们将微流体共培养平台与基于多光子成像的技术相结合，以评估同一细胞中的表型细胞行为以及 FAD 荧光强度和荧光寿命。该平台将通常对两种不同细胞群进行的两种独立测定结合到一个设备中，使我们能够同时评估表型细胞行为和酶活性。我们观察到，与亲本细胞系 (LNCaP) 相比，成骨性前列腺癌细胞系 (C4-2B) 在与骨髓基质细胞 (MC3T3-E1) 共培养时，突出表型增加，总 FAD 和蛋白结合 FAD 增加。我们假设产生 ROS 的 APAO 活性增加可能是造成这些影响的原因，并发现抗氧化剂 N-乙酰半胱氨酸 (NAC) 的存在会降低这些影响。这表明骨转移部位的ROS相关信号传导机制可能与转移性前列腺癌细胞的侵袭增加相关并在其中发挥作用。使用这个组合平台进行的研究将对驱动前列腺癌转移的机制产生新的见解。
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.