Di-n-butyl-phthalate (DBP) effect on cell viability and androgenic response in normal and cancer epithelial cells (LNCaP)

• Grant number: 13/10027-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2013
• End date: December 31, 2013
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Wellerson Rodrigo Scarano
• Grantee: Andre Rebelo Peixoto
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Di-n-butyl phthalate (DBP) - commonly used as a plasticizer in several products, including medical devices, flexible plastics and cosmetic - has anti-androgenic effect, altering signaling pathways androgen-dependent and / or independent during development of the male reproductive system. Given the worldwide problem with respect to the degradation of plastics and their dispersion in the environment, and in the earlier study where we evaluate the carcinogenic potential of DBP in the prostate of animals exposed in the peri-and postnatal care, this study aims to investigate whether treatment with DBP alters the expression of proteins involved in the processes of proliferation, stress and cell death in cultured prostatic cells. The treatment will be conducted in normal human prostatic epithelial cells immortalized (strain: RWPE-1) or cells derived from androgen dependent carcinomas (strain: LNCaP). Cells (RWPE-1 and LNCaP) will be maintained in culture medium specific and will be expanded and treated in incubator with 5% CO2 humidified atmosphere at 37 ° C. For exposure to DBP, the dose to be used shall be selected after cell viability test and determination of IC50. After selecting the optimal concentration for treatment, cells will be incubated with the specific medium containing DBP dissolved in DMSO vehicle (0.1%) for a 96 hours period. After exposure, the cells will be processed for the extraction of proteins destined for Western Blot to assess expression of the following proteins: androgen receptor (AR), PCNA, MAPK, AKT, PAR4, which are related to cellular pathways proliferation, stress and cell death. (AU)