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Prevention of cell death in LNCaP cells by overexpression of clusterin / 北京大学学报(医学版)
Article en Zh | WPRIM | ID: wpr-678863
Biblioteca responsable: WPRO
ABSTRACT
Objective: To evaluate the effect of clusterin with and without leader sequence on overexpression preventing apoptosis in human prostate LNCaP cells. Methods:The plasmid pIRES2 EGFP was used to generate the clusterin expression constructs with full length or without the leader sequence (designated as pIRES2 EGFP/cluac, pIRES2 EGFP/clubc, respectively). Western blot analysis was employed to compare clusterin expression levels in the lysis and supernatant fluid of clusterin transfected LNCaP cells in vitro . The distribution of different functional domains of clusterin in cells was detected with Immunocytochemical staining. The clusterin's protective role of Na 2SeO 3 induced apoptosis in LNCaP cells was examined by flow cytometry (FCM) and fluorescence microscope. Results: Clusterin expression was detected in the lysis and supernatant fluid of pIRES2 EGFP/cluac transfected LNCaP cells, while clusterin was found only in lysis liquid of pIRES2 EGFP/clubc transfected LNCaP cells, but not found in their supernatant fluid. The distribution of cluserin in the plasm of pIRES2 EGFP/cluac transfected cells was aggregative, and on the other hand, clusterin distributed dispersedly in pIRES2 EGFP/clubc transfected cells. Its anti apoptotic property in LNCaP cells was proved by FCM and fluorescence microscope.Conclusion: It is apparent that clusterin plays an important role in preventing apoptosis in prostate cancer, and the presence of the leader sequence is necessary for clusterin's anti apoptotic function.
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Texto completo: 1 Base de datos: WPRIM Idioma: Zh Revista: Journal of Peking University(Health Sciences) Año: 2004 Tipo del documento: Article
Texto completo: 1 Base de datos: WPRIM Idioma: Zh Revista: Journal of Peking University(Health Sciences) Año: 2004 Tipo del documento: Article