Characterization of the cellular effects of the mono-n-butyl phthalate (MBP) mediated by non-genomic estrogen pathway activated by GPR30 in human prostate tumor cells

• Grant number: 14/09191-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: September 01, 2014
• End date: February 29, 2016
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• 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

The mono-n-butyl phthalate (MBP) - active metabolite of di-n-butyl phthalate, a plasticizer commonly used in many products, including medical devices, cosmetics and flexible plastics - has estrogenic effect, changing the estrogen dependent and/or independent signaling pathways during development of the male reproductive system. Recently, it is believed that the effects of endocrine disruptors, such as DBP, to be mediated by G- coupled (GPR30) non-classical estrogen receptor protein, but there are still data showing this correlation. This receiver would be responsible for nongenomic estrogenic effects of estrogens. Given the global issues regarding the degradation of plastics and their dispersion in the environment, and before a previous study where we evaluate the carcinogenic potential of DBP in the prostate of animals exposed during the peri - and postnatal, this study has aimed to correlate the action of mono-butyl phthalate (MBP) on cells of human prostate androgen-dependent (LNCaP) and their possible effects on the verge of survival and cancer cell death, seeking to establish the correlation of effects with mechanisms of action mediated by non-classical estrogen receptor GPR30. The treatment will be carried out in human prostatic epithelial cells derived from adenocarcinomas of androgen dependent (strain:LNCaP). The cells are maintained in culture medium specific and will be expanded in the greenhouse and treated with 5% CO2 humidified atmosphere at 37°C. For exposure to MBP, the dose to be used will be selected after the MTT assay. Concomitant treatment with MBP cells will be exposed to the antagonist of GPR30 (G15). After selecting the optimal concentration for treatment, the cells are incubated with the medium containing the specific vehicle dissolved in DMSO (0.1%) for a period of 12 to 72 hours MBP. After exposure, the cells are processed for extraction of RNA and proteins, techniques aimed at Chain Reaction Real Time Polymerase after reverse transcription (RT-qPCR) and western blot. After the evaluation of gene expression of the following proteins: GPR30, EGFR, AKT, 1 MAPK (ERK2), 3 MAPK (ERK1) - ± PKC, PI3K, and c- Src, which are related to the way cell proliferation, cell death and non- genomic estrogenic effects; will perform the evaluation by western blotting of proteins that show altered gene expression. (AU)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

PEIXOTO, Andre Rebelo. Characterization of the cellular effects of the mono-N-butyl phthalate (MBP) mediated by non-genomic estrogen pathway activated by GPR30 in human prostate tumor cells. 2016. Master's Dissertation - Universidade Estadual Paulista (Unesp). Instituto de Biociências. Botucatu Botucatu.