Localization and function of the ABCC3 transporter in the prostate gland: androgen regulation and role in the protection against carcinogens

• Grant number: 15/17791-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2015
• End date: September 30, 2016
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Hernandes Faustino de Carvalho
• Grantee: André Luiz da Silva Amaral Junior
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 09/16150-6 - Androgen regulation, sinalization and cellular interactions in prostate development, physiology and regression, AP.TEM

Abstract

The use of surgical or chemical castration in the treatment of advanced prostate cancer is based on the depletion of the androgenic stimulus to keep the proliferative state of epithelial cells. Employing surgical castration and DNA microarrays, we found increased expression of the Abcc3 transporter/pump and determined that its ABCC3 product is located in the baso-lateral portion of the epithelial cells. This implies on the vectorial transport of pumped substrates toward the stroma and not along with the secretory product to the lumen. The ABCC3 transporter belogs to the ABC-type transportes and confers multidrug resistance. Many of the transporter substrates are chemoterapic drugs. In this project we seek at determining some aspcts of ABCC3 functioning and possible peculiarities of its expression and location in the prostate, in comparison with other organs. To characterize the functionality of the ABCC3 transporter, we will use two in vitro models, using the RWPE-1 prostate cells, which we showed to express the human ortholog. In the first model, the cell layer grown on Matrigel in a transwell chambers will be used to determine the vectorial transport of fluorescent drugs, in comparison with MDCK cells, derived from the kidney epithelium, given that, in this organ, the ABCC3 localizes in the apical domain of the plasma membrane. In the second model, spheroids formed by either cell type (RWPE-1 and MDCK) will be observed by time lapse in the confocal microscope to determine the luminal accumulation of fluorescent substrates in the latter and elimination of the drug in the former. Doxorubicin will be used as fluorescent substrate. Finally, we will compare the timing for drug elimination by the prostate gland and some excretory organs (pancreas, kidney, liver and intestine) in in vivo experiments. With this we aim at demonstration of a cooperation between the prostate and other organs to spare the sperm cell from the exposure to toxic compounds.