The Invasion, Migration And Proliferation Capability Of T24 Human Bladder Cancer Cell Line Influenced By RhoGDI2 Gene And Human Umbilical Vein Endothelial Cells

Objective:The overall aim of this thesis was to study the capability of T24 human bladder cancer cell line influenced by RhoGTP GDP enzyme inhibitor 2 (rho GDP dissociation inhibitor 2, RhoGDI2) and the capability of T24 human bladder cancer cell line influenced by RhoGDI2 in the three-dimensional co-culture model with human umbilical vein endothelial cells.Method:The overexpression RhoGDI2 gene of containing GFP lentiviral vector was designed and synthesized. RhoGDI2 overexpressing T24 cells was named LV-RhoGDI2-GFP/T24 cell line. GFP-load T24 cell line was named LV-CONTROL-GFP/T24 cell line. The RhoGDI2 protein expressing in LV-RhoGDI2-GFP/T24, LV-CONTROL-GFP/T24, T24 cells was verified by western blotting.The invasive ability, migration ability and proliferation ability of the three T24 human bladder cancer cell lines was detected by matrigel, transwell and CCK-8 reagent, respectively.The T24 cells, LV-RhoGDI2-GFP/T24 cells and the LV-CONTROL-GFP/ T24 cells were co-cultured with human umbilical vein endothelial cells in transwell, respectively. After 24 hours co-culture, The invasive ability, migration ability and proliferation ability of the three T24 human bladder cancer cell lines was detected by matrigel, transwell model and CCK-8 reagent, respectively.Result:The lentiviral vector containing GFP and overexpression RhoGDI2 gene was successfully constructed. The RhoGDI2 protein expression in the three T24 human bladder cancer cell lines was verified by western blotting. The RhoGDI2 protein expression of the LV-RhoGDI2-GFP/T24 cell line was significantly higher than the LV-CONTROL-GFP/T24 cell line and the T24 cell line(P<0.05=0.00). The RhoGDI2 protein expression was no significant difference between the LV-CONTROL-GFP/T24 cell line and the T24 cell line (P>0.05=0.31).In non-co-culture model, the number of invasive cells in transwell contained with matrigel of the LV-RhoGDI2-GFP/T24 cell line was significantly less than the LV-CONTROL-GFP/T24 cell line and the T24 cell line (P<0.05=0.00). The number of migrating cells in the LV-RhoGDI2-GFP/ T24 cell line was significantly less than the LV-CONTROL-GFP/T24 cell line and the T24 cell line in transwell (P<0.05=0.00). The proliferation capability of the LV-RhoGDI2-GFP/T24 cell line was significantly less than the LV-CONTROL-GFP/T24 cell line and the T24 cell line (P<0.05=0.00).In co-culture model, the number of invasive cells in transwell with matrigel of the LV-RhoGDI2-GFP/T24 cell line was significantly less than the LV-CONTROL-GFP/T24 cell line and the T24 cell line ((P<0.05=0.00). The number of migrating cells in the LV-RhoGDI2-GFP/T24 cell line was significantly less than the LV-CONTROL-GFP/T24 cell line and T24 cell line in transwell (P<0.05=0.00). The proliferation capability of the LV-RhoGDI2-GFP/T24 cell line was significantly less than the LV-CONTROL-GFP/T24 cell line and the T24 cell line (P<0.05=0.00)The co-culture model contained the human umbilical vein endothelial cells and bladder cancer cells was established by transwell, then the invasion, migration and proliferation capability of T24 human bladder cancer cell line were affected by Human umbilical vein endothelial cells were observed. The number of invasive cells of the LV-RhoGDI2-GFP/T24 cell line in non-co-cultured group and the number of invasive cells of the LV-RhoGDI2-GFP/T24 cell line in co-cultured group was no difference (,P> 0.05=0.33). However, the number of invasive cells of the LV-CONTROL-GFP/ T24 cell line in non-co-culture group was significantly less than the LV-CONTROL-GFP/T24 cell line in co-cultured group (P<0.05=0.00). The number of invasive cells of the T24 cell line in non-co-culture group was significantly less than the T24 cell line in co-cultured group (P<0.05=0.00) The number of migration cells of the LV-RhoGDI2-GFP/T24 cell line in non-co-cultured group and the number of migration cells of the LV-RhoGDI2-GFP/T24 cell line in co-cultured group was no difference (P> 0.05=0.94). However, the number of migration cells of the LV-CONTROL-GFP /T24 cell line in non-co-culture group was significantly less than the number of migration cells of the LV-CONTROL-GFP/T24 cell line in co-cultured group (P<0.05=0.00). The number of migration cells of the T24 cell line in non-co-culture group was significantly less than the number of migration cells of the T24 cell line in co-cultured group (P<0.05=0.00). The proliferation capability of the LV-RhoGDI2-GFP/T24 cell line in non-co-cultured group and the proliferation capability of the LV-RhoGDI2-GFP/T24 cell line in co-cultured group was no difference (P>0.05). However, the proliferation capability of the LV-CONTROL-GFP/T24 cell line in non-co-culture group was significantly less than the proliferation capability of the LV-CONTROL-GFP/T24 cell line in co-cultured group (P<0.05).The proliferation capability of the T24 cell line in non-co-culture group was significantly less than the T24 cell line in co-cultured group (P<0.05)Conclusion:The invasion, migration and proliferation capability of T24 human bladder cancer cell line were suppressed by RhoGDI2 gene. The invasion, migration and proliferation capability of T24 human bladder cancer cell line were promoted by Human umbilical vein endothelial cells. The promotion from human umbilical vein endothelial cell to T24 bladder carcinoma cells in invasion, migration and proliferation were interfered by RhoGDI2 gene.