HepaCAM Can Induce The Apoptosis Of Bladder Cancer Cells Via SMAD2/3/caspase Pathway

PART ONECORRELATION OF THE PROTEIN EXPRESSION OF HEPACAM AND P-SMAD2/3 IN BLADDER CANCERObjective:To explore the correlation of protein expression of hepaCAM and p-SMAD2/3 in bladder cancer and the relationship between hepaCAM or p-mTOR and various clinicopathological parameters.Methods:Immunohistochemistry staining (IHC) was used to measure the protein level of hepaCAM and p-SMAD2/3 in 51 bladder cancer tissues and 22 adjacent tissues. Disease was Ta-T1 in 13 patients, T2-T4 in 38, G1-G2 in 40, G3-G4 in 11, primary in 41 and recurrent in 10.Results:HepaCAM protein was significantly lower, and p-SMAD2/3 protein was remarkably higher in bladder cancer compared to adjacent tissues(P<0.05). Spearman correlation analysis showed the decrease of hepaCAM level was associated with the increase of p-SMAD2/3 level (r=-0.712/-0.724, P=0.008/0.011). Neither hepaCAM or p-SMAD2/3 was correlated with any of the clinicopathological parameters.Conclusion:The low expression of hepaCAM and high expression of p-SMAD2/3 are closely correlated with occurrence and development of bladder cancer, which laid a foundation for further study the mechanism of hepaCAM involving in inducing apoptosis of bladder carcinoma.PART TWOHEPACAM CAN INDUCE THE APOPTOSIS OF BLADDER CANCER CELLS VIA SMAD2/3/CASPASE PATHWAYObjective:To study the effects and molecular mechanisms of hepaCAM in inducing the apoptosis of bladder cancer cells in vitro.Methods:The flow cytometry (FCM) and Hoechst 33258 kit were used to observe the effect of hepaCAM on apoptosis of bladder cancer cells. The expression of hepaCAM and SMAD2/3/caspase pathway molecules were determined by western blot by using of hepaCAM overexpression adenovirus, caspase inhibitor (V-ZAD-FMK) and SMAD2/3 activator (TGF?1). We detected the protein expression levels of p-SMAD2/3 between nuclear and cytosol protein in bladder cancer cells BIU-87 andT24 by immunofluorescence staining and Western blot after overexpression of hepaCAM.Results:Overexpression of hepaCAM in bladder cancer cells accelerated the cell apoptosis by using of FCM and Hoechst 33258 kit. Over-expression of hepaCAM activated caspase-3/8/9, down-regulated PARP and p-SMAD2/3. Treatment with caspase pathway inhibitor (Z-VAD-FMK), compared with Z-VAD-FMK group, hepaCAM+Z-VAD-FMK group down-regulated procaspase-3/8/9 and PARP. Treatment with p-SMAD2/3 activator (TGF-?1), compared with TGF-?1 group, hepaCAM+TGF-?1 group down-regulated p-SMAD2/3, procaspase-3/8/9 and PARP. Overexpression of hepaCAM prevented the p-SMAD2/3 translocation from the cytoplasm to the nucleus in bladder cancer cell BIU-87 and T24.Conclusion:HepaCAM can induce the apoptosis of bladder cancer cells via SMAD2/3/caspase pathway.