Effect Of Nuclear Factor-Kappa B "Decoy Strategy" On Proliferation And Apoptosis Of Neuroblastoma Cell

Object To study the effect of nuclear factor kappaB (NF-κB) circular dumbbell decoyoligodeoxynucleotides (CD-ODN) transfection on the activation of NF-κB andproliferation and apoptosis of human neuroblastoma cell line SK-N-SH，and to explorepotential targets for neuroblastoma gene therapy.Methods CD-ODN was transfected into human neuroblastoma cell line SK-N-SH byelectrotransformation, and blank SK-N-SH cells were used as control groups. NF-κBnuclear translocation was studied with laser confocal microsopy. Enzyme-linkedimmunoassay (ELISA) were used to detect NF-κB DNA binding activity. Cell proliferationability was measured by MTT. Flow cytometry（FCM） was applied to analyze theapoptosis of SK-N-SH. Survivin expression was detected by western blotting. All testswere repeated for three times. The data were analyzed by statistics.Results It was confirmed by laser confocal microsopy that the phenomenon of NF-κBnuclear translocation was less obvious in SK-N-SH transfected CD-ODN. Enzyme-linkedimmunoassay (ELISA) showed the obsorbane of SK-N-SH transfected CD-ODNsignificantly decreased, compared with those of control groups,at24th,48th, and72thhours（p<0.05）.Cell proliferation was determined with MTT assay,and lower rates of MTTreduction was observed in SK-N-SH transfected CD-ODN groups than control groups, andthe cell growth inhibition rate was12%,29%and42%, respectively (p <0.05) at24th,48th, and72th hours.Flow cytometry Annexin V/PI double-staining assay showedapoptosis rates of SK-N-SH transfected CD-ODN and control groups were (23.1±0.71)%and (15.84±0.36)%(p<0.05), respectively. Early apoptosis rate was increased intransfected CD-ODN groups. Western blotting analysis showed that survivin expressionwas decreased in SK-N-SH cells treated with CD-ODN compared with the control groups. Conclusion Our results showed NF-κB circular dumbbell decoy oligodeoxynucleotides(CD-ODN) can efficiently suppress DNA binding activity of NF-κB, inhibit proliferationability of human neuroblastoma cells and may induce apoptosis of human neuroblastomacells.These experimental results indicate the CD-ODN may inhibit the occurrence anddevelopment of tumor by blocking the NF-κB signaling pathway, which may be providetheoretical basis for the neuroblastoma gene therapy.