Small-interfering-RNA-mediated silencing of human glutamate dehydrogenase induces apoptosis in neuroblastoma cells.

Abstract

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.

• Authors: Myung-Min Choi, Eun-A Kim, Jae-Wan Huh, Soo Young Choi, Sung-Woo Cho, Seung-Ju Yang
• Journal: Biotechnology and applied biochemistry (Biotechnol Appl Biochem) Vol. 51 Issue Pt 2 Pg. 107-10 (Oct 2008) ISSN: 1470-8744 [Electronic] United States
• PMID: 18248328 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Messenger
• RNA, Small Interfering
• Glutamate Dehydrogenase

Topics

• Apoptosis (drug effects)
• DNA Fragmentation (drug effects)
• Gene Expression Profiling
• Gene Silencing (drug effects)
• Genetic Vectors (genetics)
• Glutamate Dehydrogenase (biosynthesis, deficiency, genetics)
• Humans
• Immunoblotting
• Neuroblastoma (metabolism, pathology)
• RNA, Messenger (genetics, metabolism)
• RNA, Small Interfering (pharmacology)
• Reverse Transcriptase Polymerase Chain Reaction
• Transfection