Microenvironment of Prostatic Adenocarcinoma: in vivo and in vitro effect of tempol on inflammatory markers, oxidative stress, proliferation and miRNAs expression

Abstract

Prostate Cancer is the second leading cause of Cancer deaths worldwide. In Brazil it's the second most common Cancer, behind only Non-Melanoma Skin Cancer. Of the Prostate Cancer pathogenesis aspects, inflammation is an important axis for its development and progression, leading to the repeated process of tissue damage and regeneration that creates an unbalanced microenvironment of inflammatory factors and accumulation of free radicals. Tempol, a mimetic agent of Superoxide Dismutase (SOD), has been pointed to as promising coadjuvant treatment in several types of Cancer, with direct effects on the inflammatory process and oxidative stress in cells. However, studies evaluating their actions in the prostate are scarce and contradictory. The aim of the present study is to evaluate the action of Tempol, in vitro and in vivo, on the microenvironment of prostate adenocarcinoma through the analysis of inflammatory parameters, oxidative stress, tissue proliferation and miRNA expression. In vitro, the response of the cell lines LnCaP and PC3 will be analyzed after Tempol treatment, considering the aspect of dependence and androgen independence of these cells, respectively. In vivo, the response of the TRAMP (mouse transgenic mouse prostate adenocarcinoma) will be verified in two distinct stages of disease progression: 8-12 and 16-20 weeks, treated with 50 mg/kg of Tempol dose during four weeks of daily treatment. At the end of the experimental treatments, all the material will be processed for morphological analysis, gene expression (quantitative RT-PCR), Western Blotting, immunohistochemistry and TUNEL. Furthermore, based on the target genes to the pathways of inflammation and oxidative stress, it is intended to bioinformatically identify possible miRNAs of interest and to verify if they are altered with the treatment by analyzing the expression profile of these small markers in the lineages of tumor cells (PC3 and LnCaP), as well as in the prostatic tissue and in the plasma of the TRAMP model. (AU)