Therapeutic potential of BAK gene silencing in aluminum induced neural cell degeneration.
Previous studies have demonstrated robust BAK gene silencing via RNA interference (RNAi). To investigate whether BAK RNAi may serve as a co-therapeutic agent in neural cell death, we herein established a cell degeneration model using a human neuroblastoma cell line (SH-SY5Y) treated by aluminum (Al). Combining cell viability assays and expression analyses by QRT (quantitative real-time)-PCR and immunocytochemistry, we selected and validated the optimal small interfering RNA (siRNA) from three candidate siRNAs for the BAK gene. Our data identified siRNA1 as the most effective siRNA; the optimal concentration of the transfection agent was 10nM and the optimal incubation period was 24h. The transfection and knockdown efficiency was 93% and 58%, respectively, which closely correlated with the BAK protein expression. SH-SY5Y cells with BAK knockdown showed a clear resistance against cell death and Al-induced apoptosis. These results indicate that genetic inactivation of BAK could be an effective strategy in delaying the onset of apoptosis in Al-treated cells, and exemplify the therapeutic potential of RNAi-based methods for the treatment of neural cell degeneration.
Zhang
Q L
QL
Shanxi Medical University, Taiyuan, Shanxi 030001, China.
Niu
Q
Q
Shi
Y T
YT
Niu
P Y
PY
Liu
C Y
CY
Zhang
L
L
Zhang
C
C
D000535:Aluminum / Q000633:toxicity*; D017209:Apoptosis; D045744:Cell Line, Tumor; D002470:Cell Survival; D006801:Humans; D046529:Microscopy, Electron, Transmission; D009410:Nerve Degeneration / Q000139:chemically induced* / Q000473:pathology / Q000628:therapy*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000235:genetics*
D034741:RNA, Small Interfering; D050998:bcl-2 Homologous Antagonist-Killer Protein; D000535:Aluminum