Morphological and molecular study of the migration potential of prostate tumor cells exposed to extracellular matrix components

Abstract

The prostate is a gland attached to the male genital system , found only in mammals, whose main function is to produce part of the seminal fluid. The prostatic lesions arise most commonly in middle-aged men and prostate cancer (PCa) is the most diagnosed cancer and the second leading cause of cancer deaths among men in America and in Western Europe. Interactions between epithelial tissue and its surrounding stroma are responsible for maintaining the physiological function of the organ. These interactions provide proliferative and migratory constraints that define position and function of cells in the prostate tissue and are mediated by growth factors and extracellular matrix components. During cancer development, transformed cells lose these constraints, while the stroma adapts to support the "function" of the tumor. Recent findings indicate that human tumors have deregulated expression of microRNAs, molecules considered new oncogenes or tumor suppressors, used as biomarkers for diagnosis, staging and prognosis of the tumor. It is known that tumor cells acquire invasive potential migratory phenotype associated with increased expression of genes involved in cell motility, allowing these cells to respond to signals from the tumor microenvironment. Thus, studying the mechanisms and the expression of molecules involved in cell migration process appears to be highly relevant. Thus, this project aims to evaluate some parameters that determine the migration potential of human prostatic tumor cells cultured in medium containing fibronectin, collagen I and Matrigel, cultured alone or in co-culture with human fibroblasts. For this, metastatic prostate tumor cells, LNCaP and PC-3, will be exposed to different extracellular matrix components and then will be realized cell viability assays, wound healing, study of the activity and the expression of matrix metalloproteinases -2 and -9, gene expression analysis of microRNAs 21, 29b, 34a, 125b, 145, 221 and 222 and analysis of gene and protein expression of the growth factor CTGF. As a control group, the same tests will be carried out with normal immortalized prostate epithelial cells RWPE-1. (AU)