This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Animal and human population studies support the reproductive carcinogenicity of Butadiene and its metabolite BDO2. However,the toxicity and carcinogenicity of BDO2 on reproductive tissue such as the prostate or prostate cell culture has not been fully investigated. Previously we reported that one metabolite of the environmental carcinogen 1,3- Butadiene (BDO2) is more cytotoxic to androgen receptor negative (AR-) DU-145, prostate cells as compared to the androgen receptor positive (AR+) LNCaP prostate cells.
Our aim for this study was to determine if the BDO2 induce cytotoxicity in prostate cells was influenced by the presence of a functional AR. To test this question DU-145-AR- cells were transiently transfected with an AR expression plasmid (pCMVhAR), them treated with BDO2 for 24h. The presence and function of AR was determined using QRT-PCR, western blot analysis, immunocytochemistry and PSA secretion. Both western blot and immunocytochemistry analysis has demonstrated that we successfully transfected the DU-145 cells with AR protein. The AR was primarily localized to the cytoplasm and was translated into protein. Treatment of DU-145 transfected cell with BDO2 show decreased cytotoxicity as compared to untransfected DU145 cells. Functional analysis of transfected cells revealed that mRNA for AR responsive gene, and PSA, was up regulated after BDO2 treatment. Our data demonstrate that transfected of AR into the DU-145 cells decrease their sensitivity to BDO2 and restored AR transcriptional activity.
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