Bax gene silencing: a potential intervention in aluminum-induced neural cell death.
There is a vast amount of evidence indicating that Bax plays a major role in the development, maintenance, and survival of neurons and neuron-supporting cells such as glial cells. The high potency of Bax small interfering RNA (siRNA), as shown by many experimental studies, makes it a rational candidate as a co-therapeutic agent in apoptotic cell death. To investigate whether Bax RNA interference (RNAi) may serve as a potential intervention in neural cell death induced by aluminum, we herein established aluminum (Al)-treated gliatoma (C6) cells as a model for evaluating neurotoxic injury on normal glia. Using the cell model, we undertook a different approach by inducing glial cell death with Al and then using Bax gene RNAi to suppress glial cell death. Combining cell viability assays and expression analyses by quantitative real-time PCR (qRT-PCR) and immunocytochemistry, we selected and validated the optimal siRNA from 3 candidate siRNAs for the Bax gene. Sequenced reduction of neural cell death was determined with flow cytometry. Our data identified siRNA1 as the most effective siRNA. The optimal concentration of the transfection agent was 20 nM and the optimal incubation period was 72 h. The transfection and knockdown efficiencies were 95 percent and 62 percent, respectively, which closely correlated with Bax protein expression and also the cell apoptosis intervention. Taken together, Bax is essential for apoptosis induced by aluminum. Inactivation of the Bax gene could be an effective strategy for delaying the onset of apoptosis induced by Al. Our results reveal promising therapeutic potential for Bax gene silencing in Al-induced neurodegeneration.
Zhang
Q L
QL
Department of Occupational Health, School of Public Health, Shanxi Medical University, Taiyuan, China.
Niu
Q
Q
Niu
P Y
PY
Shi
Y T
YT
Liu
C Y
CY
Di Gioacchino
M
M
Zhang
L
L
Zhang
C
C
Braga
M
M
D000535:Aluminum / Q000633:toxicity*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001483:Base Sequence; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D015316:Genetic Therapy; D046529:Microscopy, Electron, Transmission; D009336:Necrosis; D009410:Nerve Degeneration / Q000235:genetics / Q000473:pathology / Q000628:therapy; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D014162:Transfection; D051028:bcl-2-Associated X Protein / Q000037:antagonists & inhibitors* / Q000235:genetics*
C494828:Bax protein, rat; D017931:DNA Primers; D034741:RNA, Small Interfering; D051028:bcl-2-Associated X Protein; D000535:Aluminum