Adenovirus-mediated delivery of bcl-2 gene attenuates cisplatin-induced degeneration of cultured spiral ganglion cells
10.3760/cma.j.issn.1673-0860.2009.11.011
- VernacularTitle:腺病毒介导bcl-2基因转染对顺铂所致大鼠螺旋神经节细胞损伤的拮抗作用
- Author:
Guo-Peng WANG
1
;
Jing XIE
;
Ying-Peng LIU
;
Hai-Hui LUO
;
Hai-Tao LU
;
Ji-Hua DONG
;
Shu-Sheng GONG
Author Information
1. 华中科技大学同济医学院附属协和医院
- Keywords:
Adenoviridae;
Transfection;
Spiral ganglion;
Genes;
bel-2;
Cisplatin;
Rats;
Sprague-Dawley
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2009;44(11):930-934
- CountryChina
- Language:Chinese
-
Abstract:
Objective To assess the protection against cisplatin-induced ototoxicity by adenovirus-mediated overexpression of the bcl-2 gene in cultured spiral ganglion cells(SGC).Methods SGC from P3 rats were cultured in vitro and exposed to adenovirus vector carrying green fluorescent protein gene(Ad-GFP),followed by immunocytochemical analysis for expression of the neuron-specific marker Neurofilament200(NF200)and detection under laser scanning confocal fluorescence microscope. Then, SGC were transduced by Ad-bel-2 and the expression of human bcl-2 protein was evaluated by Western Blot. Finally, the cultures of SGC were divided into 4 groups: the group of Ad-bcl-2 transfection followed by cisplatin treatment, the group of Ad-GFP transfection followed by cisplatin treatment, the group of cisplatin treatmentonly and the untreated group. Cisplatin worked for 48 hours at a concentration of 2μg/ml. Outcome measures included survival humber of SGC and longest neurite length by using ImageJ software. Results SGC were cultured successfully in vitro and transfected by adenovirus vector safely and efficiently. By Western Blot, human bcl-2 protein was expressed in the group after exposure to Ad-bcl-2,but not in the Ad-GFP transfected SGC. Cisplatin exposure resulted in shrinking of neuritis and pyknosis of cell body, even cell death. Expression of bel-2 in the SGC provided a significant level of protection against cisplatin-induced SGC degeneration. Conclusions Our results suggest that SGC can be transduced by adenovirus vector safely and efficiently in vitro. Adenovirus-mediated delivery of the bcl-2 gene attenuates cisplatin-induced SGC degeneration.
