Experimental Investigation On Invasion And Metastasis Potentials Of Human Bladder Cancer Cells T24 Under There-Dimensional Culture Modal

Objective:To establish bladder cancer cell T24 3D culture model; to investigate the effects of microenvironment on morphological characteristics of bladder cancer cell T24 and to investigate the expression of Eph A2,MMPs and focal adhesion molecules E-cadherin under 3D cell culture condition, and to study the mechanism of bladder cancer T24 adhesion, invasion and metastasis in three-dimensional culture model.Method: Firstly bladder cancer T24 cells were cultured in 2D cell culture condition, the bladder cancer T24 cells in good condition were obtained with method of trypsinization.T24 cells were cultured in completely melt Matrigel which was diluted with RPMI1640 to different proportion, some T24 cells were seeded onto the surface of concretionary Matrigel, and we also have T24 cells culture in completely melt Matrigel directly. Thus to explore and establish bladder cancer cell T24 3D culture model.When the model was established, we observed the morphological changes of cells with microscope and investigated the expression of Eph A2, MMPs(MMP2 and MMP9)and E-cadherin protein in T24 cells under 3D and 2D culture model with the method of West-blotting. The cell invasion ability under these two different cell culture modals was detected by using Transwell invasion assay.Result:The morphological changes of T24 cells under different culture conditions? T24 cells cultured in the diluted Matrigel cannot establish 3D culture model. We found cells seeded onto the surface of Matrigel can form part three-dimensional cell clones. When cells culture in completely melt Matrigel directly, non-polarity spherical cell clones were cultivated with the extending of cultured time, and the surface of cell clones exhibited some filopodia gradually. The 3D cell culture model was established this way.Expression levels of Eph A2, MMPs( MMP2 and MMP9) and E-cadherin protein in T24 cells under different culture modal? The Expression levels of Eph A2, MMPs(MMP2 and MMP9)and E-cadherin protein in T24 cells in 3D culture modal were higher than 2D culture modal(P<0.05).? Similarly, The cell invasion ability under 3D cell culture modal has stronger invasiveness(P<0.05).Conclusion:1. bladder cancer T24 cells 3D cell culture modal can be established by using extracellular matrix- Matrigel.2. under 3D cell culture condition, enhancing the expression of Eph A2,MMPs(MMP2 and MMP9) and adhesion molecules E-cadherin possibly be the mechanism of increasing the adhesion and invasion potentials of T24 cells.