Reciprocal changes in gene expression profiles of cocultured breast epithelial cells and primary fibroblasts

Abstract

The importance of epithelial-stroma interaction in normal breast development and tumor progression has been recognized. To identify genes that were regulated by these reciprocal interactions, we cocultured a nonmalignant (MCF10A) and a breast cancer derived (MDA-MB231) basal cell lines, with fibroblasts isolated from breast benign-disease adjacent tissues (NAF) or with carcinoma-associated fibroblasts (CAF), in a transwell system. Gene expression profiles of each coculture pair were compared with the correspondent monocultures, using a customized microarray. Contrariwise to large alterations in epithelial cells genomic profiles, fibroblasts were less affected. In MDA-MB231 highly represented genes downregulated by CAF derived factors coded for proteins important for the specificity of vectorial transport between ER and golgi, possibly affecting cell polarity whereas the response of MCF10A comprised an induction of genes coding for stress responsive proteins, representing a prosurvival effect. While NAF downregulated genes encoding proteins associated to glycolipid and fatty acid biosynthesis in MDA-MB231, potentially affecting membrane biogenesis, in MCF10A, genes critical for growth control and adhesion were altered. NAFs responded to coculture with MDA-MB231 by a decrease in the expression of genes induced by TGFbeta1 and associated to motility. However, there was little change in NAFs gene expression profile influenced by MCF10A. CAFs responded to the presence of both epithelial cells inducing genes implicated in cell proliferation. Our data indicate that interactions between breast fibroblasts and basal epithelial cells resulted in alterations in the genomic profiles of both cell types which may help to clarify some aspects of this heterotypic signaling.

Carcinoma-associated fibroblasts: orchestrating the composition of malignancy

“…Several studies have shed light on these questions and contributed to a stunning realization that stromal changes and the existence of CAFlike cells can precede the formation of malignant epithelial cells. While it is clear that coculturing fibroblasts and malignant epithelial cell lines can result in reciprocal gene expression changes (Rozenchan et al 2009), characterization of tissues at high risk for carcinoma formation demonstrate that stromal changes associated with the CAF phenotype already exist and are therefore not dependent on signals from a carcinoma for their generation (Saadi et al 2010;DeFilippis et al 2014).…”

Role of cancer-associated fibroblasts in breast cancer development and prognosis

“…This indicates that paracrine and endocrine effects are at the origin of these modifications since the basement membrane is largely intact in DCIS. Using a transwell system allowing diffusible factor exchange and microarray analysis, it has been shown that 160 and 178 genes were differentially expressed between MDA-MB231 and MCF-10 cells cocultured with CAFs as compared to monocultures, respectively (Rozenchan et al, 2009). Moreover, co-injection of lethally irradiated fibroblasts or inclusion of fibroblast-conditioned medium with breast cancer cell grafts increased carcinogenesis and tumor growth (Camps et al, 1990;Noel et al, 1993).…”

Galectin-1 knockdown in carcinoma-associated fibroblasts inhibits migration and invasion of human MDA-MB-231 breast cancer cells by modulating MMP-9 expression

“…Carcinoma-associated fibroblasts (CAFs, commonly referred to as myofibroblasts), the most abundant cell type in breast cancer stroma, are becoming increasingly apparent due to its effect on promoting tumor cell growth, migration, invasion, and drug resistance [2]. Increasing evidence indicated that CAFs could induce breast epithelial cell migration and invasion by producing a plethora of chemokines, growth factors, and ECM proteins [3], and a dynamic bidirectional signaling is detected between CAFs and breast cancer cells [4,5]. However, the precise molecular mechanisms of CAFs' effect on the migration and invasion of breast cancer are still far from clear.…”