Overexpression of XIAP inhibits apoptotic cell death in an oligodendroglial cell line.
1. This study describes the use of an oligodendroglial cell line (158N) to study the protective effects of X-chromosome-linked inhibitor of apoptosis (XIAP) overexpression. 2. 158N cells were transiently transfected with either pCMV-Myc-XIAP or control pCMV-Myc vector. At 48 h post-transfection, immunoblotting and immunocytochemical staining showed robust myc-XIAP overexpression in pCMV-Myc-XIAP transfected cells relative to pCMV-Myc-transfected cells and normal 158N cells. 158N cells were treated with either 100 nm staurosporine (STS) or 300 microM dopamine (DA) and cell survival/function determined using two cell viability assays. 3. Both STS and DA treatments resulted in increased apoptotic death of pCMV-Myc transfected cells. In contrast, there was significant decrease in apoptotic cell death in cells transfected with pCMV-Myc-XIAP. Finally, XIAP overexpression was found to significantly reduce caspase-3 enzyme activity levels in response to apoptotic stimuli. 4. These results provide evidence that XIAP overexpression promotes cell survival in a non-neuronal cell type derived from the central nervous system. In addition, these data suggest that the 158N oligodendroglial cell line is a suitable tool for transient transfection studies, which is a problem frequently encountered when attempting to introduce genes of interest in cultures of primary oligodendroglia.
Jin
Ying
Y
Department of Anatomy and Neurobiology, Spinal Cord and Brain Injury Research Center, University of Kentucky Medical Center, Lexington, KY 40536-0298, USA.
McEwen
Melanie L
ML
Ghandour
M Said
MS
Springer
Joe E
JE
D017209:Apoptosis / Q000502:physiology*; D016923:Cell Death / Q000502:physiology; D002461:Cell Line, Transformed; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods; D051636:X-Linked Inhibitor of Apoptosis Protein
D011506:Proteins; D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human