Small-interfering-RNA-mediated silencing of human glutamate dehydrogenase induces apoptosis in neuroblastoma cells.
In the nervous system, GDH (glutamate dehydrogenase) is enriched in astrocytes and is important for recycling glutamate, a major excitatory neurotransmitter. The function of hGDH (human GDH) may be important in neurodegenerative diseases such as Parkinson's disease. To test the effect of decreased hGDH expression, several vector-based plasmidlinked hGDH siRNAs (small interfering RNAs) were expressed intracellularly in BE(2)C human neuroblastoma cells. Immunoblotting and reverse-transcription-PCR confirmed that expression of hGDH protein and mRNA was knocked down by co-transfection with phGDH-siRNA vectors in BE(2)C human neuroblastoma cells. TUNEL (terminal uridine deoxynucleotidyl transferase dUTP nick-end labelling) and DNA fragmentation assays 48 h after transfection of phGDH-siRNAs revealed that inhibition of hGDH expression induced cellular apoptosis and activated phospho-ERK1/2 (phospho-extracellular-signal-regulated kinase 1/2). These findings show that inhibition of hGDH expression by siRNA is related to apoptosis in neuronal cells.
Choi
Myung-Min
MM
Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine, Seoul, Korea.
Kim
Eun-A
EA
Huh
Jae-Wan
JW
Choi
Soo Young
SY
Cho
Sung-Woo
SW
Yang
Seung-Ju
SJ
D017209:Apoptosis / Q000187:drug effects*; D053938:DNA Fragmentation / Q000187:drug effects; D020869:Gene Expression Profiling; D020868:Gene Silencing / Q000187:drug effects*; D005822:Genetic Vectors / Q000235:genetics; D005969:Glutamate Dehydrogenase / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D005969:Glutamate Dehydrogenase