Effect Of Hepacam Gene On Adhesion And Its Mechanism Of Human Bladder Cancer Cell Line T24

Objective To investigate the adhesiveness and its mechanism of transfection with exogenous wild hepaCAM gene on biological behavior of the transitional cell carcinoma of bladder(TCCB) in vitro.Methods Plasmid pEGFP-N2-hepaCAM was transfected into bladder cancer cell(T24) by LipofectamineTM 2000,and cells that can express hepaCAM stably was screened out by G418(400μg/ml)。Western bolt was used to determine the expression of target of hepaCAM in stable cells。The tumor cell adhere to the vascular endothelial cell to use to research the adhesiveness of hepaCAM gene on T24 cell。Using laser confocal microscopy to detect the cytoskeleton in T24 cells。The actin content in T24 cell measured with flow cytometry.Result Acquired T24 cell clone with stable expression of hepaCAM。In vitro experiments as tumor cell adhere to the vascular endothelial cell showed the cell adhesion was apparently enhanced compared with the non-transfection and mock-vehicle group(sP<0.05).In hepaCAM group,the fluorescence of filamentous actin(F-actin) was strengthen. The structure of filamentous actin was condensed and regularity.In sham and control group,the result was opposite.The content of F-actin in hepaCAM group was apparently enhanced compared with the non-transfection and mock-vehicle groups(P<0.05).Conclusion The hepaCAM gene can enhance the adhesiveness of T24 cell by induce F-actin cytoskeleton ,and may inhibit the TCCB invasion and metastasis.