Inducing apoptosis of bladder cancer cell T24 by 5-azacytidine combined with adenovirus-hepaCAM via inhibiting p-AKT

Abstract:

Objective：To explore the effect of 5-azacytidine（azac） combined with adenovirus-hepatocyte cell adhesion molecule（ad-hepaCAM） on the apoptosis of bladder cancer cells T24 and the possible mechanisms. Methods：Bladder cancer cells T24 were treat－ed with seven different factors. Apoptosis rate of each treatment group was detected by the kits of hoechst 33258. Cell apoptotic ratio at early stage was detected by flow cytometry. The mRNA expression of hepaCAM and bcl-2 was measured by RT-PCR. The protein expression of hepaCAM，p-AKT，total-AKT and bcl-2 was measured by Western blot. Results：Higher apoptosis rate in azac combined with ad-hepaCAM group（45.21±0.06）% than in adenovirus-hepaCAM group（15.60±0.02）% and azac group（26.30±0.02）% was observed based on hoechst 33258（P=0.003，P=0.008）. Higher apoptosis rate at early stage in azac combined with ad-hepaCAM group group（16.05±0.06）% than in adenovirus-hepaCAM group（3.90±0.02）% and azac group（9.67±0.02）% was observed based on FCM（P=0.002，P=0.043）. Higher expression of hepaCAM mRNA（P=0.004，P=0.000） and lower expression of bcl-2 mRNA（P=0.026，P=0.030） in azac combined with ad-hepaCAM group than in adenovirus-hepaCAM group and azac group was observed based on RT-PCR. Higher expression of hepaCAM（P=0.003，P=0.003） and lower expression of p-AKT（P=0.001，P=0.005） and bcl-2（P=0.000，P=0.002） in azac combined with ad-hepaCAM group than in adenovirus-hepaCAM group and azac group was observed based on Western blot. Conclusion：Azac combined with ad-hepaCAM can induce the apoptosis of T24 by inhibiting p-AKT，which may provide a new therapy for bladder cancer.