PO-242 Novel analysis of cancer cell invasion using a three-dimensional cultured tissue model mimics human tissue organisation

Abstract

IntroductionPreventing cancer metastasis requires a through understanding of cancer cell invasion. Two-dimensional (2D) culture systems are typically used to analyse these processes; for instance, wound healing and transwell assays are used to evaluate cell migration, whereas the Matrigel chamber assay is used to assess cell invasion. However, these may not reflect actual events that occur in human tissue. We developed a human cell-based three-dimensional (3D) cultured tissue model that mimics human tissue and used it evaluate cell invasion in human oral squamous cell carcinoma (OSCC).Material and methodsThe 3D tissue structure consisted of five layers of normal human dermal fibroblasts along with human dermal lymphatic endothelial cell (HDLEC) tubes and was generated by the cell accumulation technique and layer-by-layer assembly using fibronectin and gelatin. OSCC cells with different lymph metastatic capacity were inoculated on the 3D tissues and their invasion through the 3D tissue structure was observed. Highly lymph metastatic SAS-LM8 and HSC-LM3 cells were established from the SAS-Venus and HSC-Venus cell lines through in vivo selection individually. Conventional methods were traswell chamber assay and Matrigel chamber assay also used for comparison.Results and discussionsThe results of invasion capacity using the 3D cultured tissue model were comparable to those obtained using conventional 2D culture-based transwell and Matrigel chamber assays. In addition, our findings using the 3D cultured tissue model are consistent with those of our previous study reporting that SAS-Venus cell migration and invasion were increased by stimulation with Wnt5b. Importantly, we clearly observed invasion of these cancer cells into the 3D tissue structure and HDLEC cells formed capillary-like extensions in the 3D structure. Moreover, we could visualised the distance from each cancer cell to HDLECs based on immunofluorescence labelling.ConclusionOur 3D cultured tissue system allows detailed analysis of cancer cell migration and invasion in an environment that mimics actual human tissue. It is expected that the expression of genes and proteins involved in these processes. It is a very useful tool for analysing a process of cancer cell invasion through a human living tissue.