Proapoptotic effects of NF-kappaB on cisplatin-induced cell death in auditory cell line.
CONCLUSIONS
The findings suggest that nuclear factor (NF)-kappaB is activated by cisplatin and plays a proapoptotic role during cell death in the auditory cell line HEI-OC1.


OBJECTIVE
Cisplatin is a very effective antineoplastic drug but in high doses it shows ototoxicity by inducing apoptosis of hair cells in the cochlea. NF-kappaB is a transcription factor regulating apoptosis in many organs and tissues. This study examined the role of NF-kappaB in the apoptotic pathway induced by cisplatin in the auditory cell line, HEI-OC1.


METHODS
Apoptotic cell death was identified by measuring caspase 3 activity and immunostaining with anti-caspase 3 antibody after cisplatin treatment (50 microM) for 24 h. To investigate the role of NF-kappaB in apoptotic cell death, HEI-OC1 cells treated with 50 microM of cisplatin were immunostained with anti-NF-kappaB (p65) antibody. Two different NF-kappaB inhibitors, Bay 11-7085 and SN-50, were co-incubated with cisplatin (50 microM) for 24 h and caspase 3 activity was assayed.


RESULTS
Immunostaining with anti-caspase 3 antibody and caspase 3 assay showed that cisplatin induced apoptosis in HEI-OC1 cells. After cisplatin treatment, NF-kappaB (p65) was activated to translocate from the cytoplasm into the nucleus. Co-treatment with NF-kappaB inhibitors reduced the cisplatin-induced apoptosis of HEI-OC1 cells.
Chung
Won-Ho
WH
Department of Otorhinolaryngology-Head and Neck Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. whchung@skku.edu
Boo
Sung Hyun
SH
Chung
Man Ki
MK
Lee
Hyun-Seok
HS
Cho
Yang-Sun
YS
Hong
Sung Hwa
SH
D000818:Animals; D000906:Antibodies / Q000032:analysis; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001692:Biological Transport; D053148:Caspase 3 / Q000276:immunology; D016923:Cell Death; D002460:Cell Line; D002945:Cisplatin / Q000494:pharmacology*; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000502:physiology*; D009570:Nitriles / Q000494:pharmacology; D009925:Organ of Corti / Q000166:cytology; D010455:Peptides / Q000494:pharmacology; D013450:Sulfones / Q000494:pharmacology
D000906:Antibodies; D000970:Antineoplastic Agents; C416282:BAY 11-7085; D016328:NF-kappa B; D009570:Nitriles; D010455:Peptides; C118258:SN50 peptide; D013450:Sulfones; D053148:Caspase 3; D002945:Cisplatin