Apoptosis of Prostate Cancer by Bax Gene Expression.
- Author:
Cheol KWAK
1
;
Ren Jie JIN
;
Ja Hyeon KU
;
Hyeon JEONG
;
Eun Sik LEE
;
Sang Eun LEE
;
Chong Wook LEE
Author Information
1. Department of Urology, Seoul National University College of Medicine, Seoul, Korea. urology@snu.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Bax protein;
Prostatic neoplasms;
Apoptosis;
Gene therapy
- MeSH:
Apoptosis*;
bcl-2-Associated X Protein;
bcl-X Protein;
Blotting, Western;
Cell Line;
Clone Cells;
DNA Fragmentation;
DNA, Complementary;
Gene Expression*;
Genetic Therapy;
Humans;
Parents;
Plasmids;
Prostate*;
Prostatic Neoplasms*;
RNA, Messenger;
Transfection
- From:Korean Journal of Urology
2003;44(9):916-923
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: To evaluate the antitumor effect of the proapoptotic Bax gene in prostate cancer cells, in vitro, using a plasmid vector expressing the human Bax gene. MATERIALS AND METHODS: cDNA of the human Bax gene, amplified by RT-PCR, was cloned to pCR@3.1. The expression of the cloned Bax (pCR3.1-Bax) was observed by RT-PCR and Western blot analyses. The efficacy of growth inhibition by the cloned Bax gene was tested, in vitro, on PC-3 and DU145 human prostate cancer cell lines using the MTT assay. Immunoblot analysis for the expressions of Bcl-2 and Bcl-xL were performed. Assays were also performed to evaluate the apoptosis, DNA fragmentation and CPP32. RESULTS: The Bax protein was expressed in the parental PC-3 cells, but not in the DU-145 cells. The expressions of Bax mRNA in the transfected PC-3 and DU-145 cells had increased by 24 hr, and those of Bax protein in the transfected PC-3 and DU-145 cells had increased by 48 and 24 hr, respectively, compared with the control cell lines. The cytotoxicity of pCR3.1-Bax on PC-3 and DU-145 cells increased significantly compared with an empty vector, pCR3.1 (p<0.05, respectively). An increased cytotoxicity of the Bax-transfected cell lines was associated with enhanced apoptosis. The Bcl-2 protein was not expressed in the transfected cells, and the levels of Bcl-xL protein expression in transfected cells were no different to those in the parenteral cells. The Bax/Bcl-xL ratio was increased by the transfection of the Bax expression vector. CONCLUSIONS: Our results show that the cloned Bax-expression plasmid vector efficiently inhibits the growth of PC-3 and DU145 human prostate cancer cell lines. These data suggest that exogenous Bax expression may have therapeutic applications in prostate cancer.
