Bak: a downstream mediator of fenretinide-induced apoptosis of SH-SY5Y neuroblastoma cells.
Unlike 13-cis-retinoic acid, the synthetic retinoid fenretinide [N-(4-hydroxyphenyl)retinamide] induces apoptosis of neuroblastoma cells by mechanisms involving retinoic acid receptors and oxidative stress. After screening a cDNA array for apoptosis-related genes, the Bcl2-related protein Bak was identified as a fenretinide-inducible gene in SH-SY5Y neuroblastoma cells, and this was confirmed by Western blotting and flow cytometry. Although fenretinide acts synergistically in vitro with chemotherapeutic drugs, these drugs did not induce Bak expression. Retinoic acid receptor antagonists did not block the induction of Bak by fenretinide. Conversely, Bak induction was blocked by the antioxidant vitamin C. Overexpression of Bak increased apoptosis in both the presence and absence of fenretinide, whereas expression of antisense Bak inhibited fenretinide-induced apoptosis. Bak expression was also induced in cells overexpressing the stress-induced transcription factor GADD153, but Bak expression was inhibited in cells expressing an antisense GADD153 construct. These results suggest that Bak is a downstream mediator of an oxidative stress pathway leading to apoptosis of SH-SY5Y neuroblastoma cells in response to fenretinide.
Lovat
Penny E
PE
Northern Institute for Cancer Research, 4th Floor Cookson Building, Medical School, University of Newcastle, Newcastle Upon Tyne NE2 4HH, United Kingdom.
Oliverio
Serafina
S
Corazzari
Marco
M
Rodolfo
Carlo
C
Ranalli
Marco
M
Goranov
Bojidar
B
Melino
Gerry
G
Redfern
Christopher P F
CP
Piacentini
Mauro
M
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D017313:Fenretinide / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis / Q000502:physiology*; D009447:Neuroblastoma / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D050998:bcl-2 Homologous Antagonist-Killer Protein
D000970:Antineoplastic Agents; C494772:BAK1 protein, human; D008565:Membrane Proteins; D050998:bcl-2 Homologous Antagonist-Killer Protein; D017313:Fenretinide