The effect of sFLT01 gene expression on cell proliferation and migration of bladder cancer cell line; 5637

Bladder cancer is the most common urinary malignancy all over the world. . The pathologic angiogenesis plays an important role in bladder cancer tumor growth and progression .VEGF is one of the primary required angiogenic factor for bladder cancer and is able to induce tumor angiogenesis and accelerate tumor growth.sFLT01 is an engineered chimeric secretory receptor/protein with the inhibitory effect on VEGF and placental growth factor(PLGF). The objective of this study is to evaluate the effect of sFLT01 over expression on proliferation and migration of 5637 bladder cancer cell line. sFLT01-His Tag-GFP sequence was designed synthesized and cloned in AAV-MCS-GFP vactor. PAAV-sFLT01-His Tag-GFP vector was transfected to 5637cell line through lipofection 2000 transfection. Then extracted mRNA was analyzed by real time PCR. Protein secretion into the conditioned medium of transfected 5637cells and HEK293T cells proved by western blotting. The effect of the condition medium 5637 cells and HEK293T cells on in vitro angiogenesis in HUVEC cells was investigated by the invitro angiogenesis assay.5637 cell migration was evaluated by scratch assay. The cytotoxic effect of the constructson 5637 cells was determined and cell proliferation assay was performed through the MTT assay.Real-Time PCR results showed significant over expression of sFLT01 in treated 5637 cells.Western blot proved sFLT01 protein secretion in conditioned media. In vitro angiogenesis assay results showed decreased potential of tube formation in conditioned medium of treated 5637 cells and HEK293T cultures. Scratch assay showed no significant difference in treated 5637 cells when compared to the control untreated cultures. MTT assay showed that sFLT01 had no cytotoxic effect on 5637 cells.