Genomics and proteomics approaches to the study of cancer-stroma interactions

Flávia C Rodrigues-Lisoni,Erica E Fukuyama,Marcos B De Carvalho,Tiago Henrique,Giovana M Polachini,Eloiza H Tajara,Pedro Michaluart,Rodrigo Ap Teixeira,Caique F Souza,Alessandra Vidotto,Bianca R Cunha,Sonia M Oliani,José V Maniglia,Juliana Carmona-Raphe,Paulo Peitl

doi:10.1186/1755-8794-3-14

Abstract

BackgroundThe development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression.MethodsThe study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells.ResultsWe observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (ARID4A, CALR, GNB2L1, RNF10, SQSTM1, USP9X) were validated by real time PCR.ConclusionsA significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.

Highlights

The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment
The spontaneously immortalized cell line of fibroblasts retained the characteristics of stromal cells and may correspond to cancer-associated fibroblasts (CAF)
We investigated the influence of soluble paracrine factors produced in vitro by stromal cells derived from an oral carcinoma and by a neoplastic epithelial cell line on proliferation and gene/protein expression

Summary

Introduction

The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression. Tumor cells may alter the surrounding stroma through direct cell contact or via the secretion of paracrine soluble factors, inducing cell differentiation or extracellular matrix modifications [9]. In it turn, stromal cells may promote cancer progression and acquisition of invasiveness [10,11,12]. As Albini and Sporn (2008) appropriately propose, the microenvironment may be more than a partner and an essential component of the cancer, and both should be considered as a functional whole [15]

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