| Summary: | Cancer is a fatal disease that cause overgrowth of cells. The spreading of these cells from a primary to secondary site is termed as cancer metastasis and this involves a very important step, extravasation. For studying this step, transwell assays are used but they pose a few problems. Furthermore, the role of chemicals, H2O2 and VEGF in promoting transendothelial migration and the location cancer cell can survive are still unravelled. Hence, this study aimed to develop a new in vitro assay to study extravasation and investigate these unknowns. For this, HUVEC-DT and 231-C3 cells were used to generate a 2-colour co-culture system. General procedures such as passaging, counting, seeding and imaging were performed. Using both this model and also in real-time, we found that cancer cells were able to disrupt cell-cell junction of the endothelial monolayer and invade into it on a cultural dish. We termed this effect of cancer cells on endothelial monolayer as endothelial breakage. Moreover, the chemicals were found to promote endothelial breakages of cancer cells. We also observed that the cancer cells attached to endothelial cells showed less cell spreading and higher apoptosis rate, compared to cancer cells that attached to the dish bottom. This could be mostly due to the endothelial cell receptors which could bind to ligands of cancer cells and provide signals that trigger apoptosis pathways. Hence, these new findings will help advance the understanding of cancer metastasis and also our co-culture model can be used in various applications such as drug screening.
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