A double-strand decoy DNA oligonucleotides of NF-κB induced the apoptosis and sensitization of hepatic cancer cells to chemotherapy

Objective To investigate a double-stranded decoy oligodeoxynucleotides (ODNs) with a specific affinity to NF-κB effectively suppressed NF-κB activity and sensitized hepatic cancer cells to chemotherapy.Methods HepG2 cells resistanting to adriamycin (HepG2/ADR) were induced step- wised,FITC-labeled decoy ODNs against NF-κB were transfected into HepG2/ADR cells with Lipofec- tAMINE~(TM)2000,Inverse fluorescent microscopand showed the localization of decoy DNA,and EMSA was performated to investigate the affinity of NF-κB.ADM (0.1 mg/L) was added,the apoptosis was ob- served by flow cytometry and TUNEL differently.The activity of caspase3 was found by quantitative as- say,the expression of bc1-2 protein was detected by western blot.Results HepG2/ADR was confirmed resisting to ADM.When decoy ODNs was transfected,after 1 hour,FITC-labeled decoy ODNs against NF-κB was detected in the nuclei of HepG2/ADM cells,and electrophoretic mobility shift assays (EM- SA) was performed to found the activation of NF-κB binding to the nucleus was inhibitd.Incubed with ADM (0.1 mg/L),it showed significant inhibitory effect on the growth of HepG2/ADM,the percentage of apoptosis was increased compare with control at 24 h,caspase3 activity was increased,bc1-2 expression was downregulated.Conclusion Our findings suggest that decoy ODNs for NF-κB could be a novel and attractive strategy to improve treatment outcome of human hepatocarcinoma,bc1-2 and caspase3 could play important role during decoy ODNs inducing apoptosis.