Abstract
Invadopodia or invasive feet, which are actin-rich membrane protrusions with matrix degradation activity formed by invasive cancer cells, are a key determinant in the malignant invasive progression of tumors and represent an important target for cancer therapies. In this work, we presented a microfluidic 3D culture device with continuous supplement of fresh media via a syringe pump. The device mimicked tumor microenvironment in vivo and could be used to assay invadopodia formation and to study the mechanism of human lung cancer invasion. With this device, we investigated the effects of epidermal growth factor (EGF) and matrix metalloproteinase (MMP) inhibitor, GM6001 on invadopodia formation by human non-small cell lung cancer cell line A549 in 3D matrix model. This device was composed of three units that were capable of achieving the assays on one control group and two experimental groups' cells, which were simultaneously pretreated with EGF or GM6001 in parallel. Immunofluorescence analysis of invadopodia formation and extracellular matrix degradation was conducted using confocal imaging system. We observed that EGF promoted invadopodia formation by A549 cells in 3D matrix and that GM6001 inhibited the process. These results demonstrated that epidermal growth factor receptor (EGFR) signaling played a significant role in invadopodia formation and related ECM degradation activity. Meanwhile, it was suggested that MMP inhibitor (GM6001) might be a powerful therapeutic agent targeting invadopodia formation in tumor invasion. This work clearly demonstrated that the microfluidic-based 3D culture device provided an applicable platform for elucidating the mechanism of cancer invasion and could be used in testing other anti-invasion agents.
Highlights
Lung cancer causes the most deaths in human beings among all cancers [1]
The blue indicator represented control group, the red indicator represented epidermal growth factor (EGF) group, while the green indicator represented GM6001/EGF group. These indicators could spread in the cell chambers of both sides via oval microchannels uniformly and in parallel without crossing, showing that the microfluidic system was practicable for the assay of the invadopodium formation in A549 lung cancer cell line with several groups
We developed a simple but effective microfluidic device to investigate the role of EGF, GM6001 in invadopodia formation in the process of invasion for A549 lung cancer cell line in an in vivo-like 3D microenvironment
Summary
Lung cancer causes the most deaths in human beings among all cancers [1]. The initial stage of cancer cell migration and invasion is the extension of cell protrusions in the direction of cell movement. The formation of these cell protrusions is usually driven by actin polymerization at the leading edge [3,4]. Many reports on invadopodia formation with mammary adenocarcinoma, oral squamous carcinoma, colon cancer, melanoma, etc., have been published [5,6,7,8,9,10,11,12,13], whereas, only one involved lung adenocarcinoma [14]
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