Identification of Molecular Distinctions Between Normal Breast-Associated Fibroblasts and Breast Cancer-Associated Fibroblasts

Abstract

Stromal fibroblasts influence the behavior of breast epithelial cells. Fibroblasts derived from normal breast (NAF) inhibit epithelial growth, whereas fibroblasts from breast carcinomas (CAF) have less growth inhibitory capacity and can promote epithelial growth. We sought to identify molecules that are differentially expressed in NAF versus CAF and potentially responsible for their different growth regulatory abilities. To determine the contribution of soluble molecules to fibroblast–epithelial interactions, NAF were grown in 3D, transwell or direct contact co-cultures with MCF10AT epithelial cells. NAF suppressed proliferation of MCF10AT in both direct contact and transwell co-cultures, but this suppression was significantly greater in direct co-cultures, indicating involvement of both soluble and contact factors. Gene expression profiling of early passage fibroblast cultures identified 420 genes that were differentially expressed in NAF versus CAF. Of the eight genes selected for validation by real-time PCR, FIBULIN 1, was overexpressed in NAF, and DICKKOPF 1, NEUREGULIN 1, PLASMINOGEN ACTIVATOR INHIBITOR 2, and TISSUE PLASMINOGEN ACTIVATOR were overexpressed in CAF. A higher expression of FIBULIN 1 in normal- than cancer-associated fibroblastic stroma was confirmed by immunohistochemistry of breast tissues. Among breast cancers, stromal expression of Fibulin 1 protein was higher in estrogen receptor α-positive cancers and low stromal expression of Fibulin 1 correlated with a higher proliferation of cancer epithelial cells. In conclusion, expression profiling of NAF and CAF cultures identified many genes with potential relevance to fibroblast–epithelial interactions in breast cancer. Furthermore, these early passage fibroblast cultures can be representative of gene expression in stromal fibroblasts in vivo.Electronic supplementary materialThe online version of this article (doi:10.1007/s12307-008-0017-0) contains supplementary material, which is available to authorized users.

Functional Heterogeneity of Cancer-Associated Fibroblasts from Human Colon Tumors Shows Specific Prognostic Gene Expression Signature

“…Thus, in breast cancer, significant differences between CAFs and normal counterparts or CAFs derived from different clinical subtypes have been described previously (21)(22)(23). In nonsmall cell lung cancer, genes relating to the TGF-b signaling pathway are differently expressed by NFs and CAFs (24).…”

Carcinoma-associated fibroblasts: orchestrating the composition of malignancy

“…To obtain clues as to the stromal pathways involved in CAF-dependent facilitation of malignant phenotypes, several studies have compared gene expression in disease-free fibroblasts and CAFs derived from various tissues (Sadlonova et al 2009;Saadi et al 2010;DeFilippis et al 2012;Berdiel-Acer et al 2014). In one of these studies conducted in breasts, the most dramatic change in expression between disease-free fibroblast and CAF populations was the repression of a cell surface protein, CD36.…”

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

“…These genes are implicated in paracrine or intracellular signaling, transcriptional regulation, ECM production and cell adhesion/migration (Bauer et al, 2010). In another study, 420 genes were found to be differentially expressed in CAFs versus NBFs (Sadlonova et al, 2009). Furthermore, 2-Dimentional (2D) gel electrophoresis analysis revealed a great variation in global protein expression between CAFs, NFs and NBFs.…”

“…These factors are used by activated fibroblasts to communicate with cancer cells as well as with other stromal cells. Microarray and proteomics analysis have revealed distinct mRNA and protein expression profiles in CAFs as compared to their adjacent normal counterparts (NFs) and normal fibroblasts isolated from mammary reduction surgery (NBFs), showing genetic and/or epigenetic alterations in these stromal fibroblasts (Allinen et al, 2004;Bauer et al, 2010;Hawsawi et al, 2008;Sadlonova et al, 2009). Consistent with this, CAFs exhibit some cancer-specific changes, including defective p53/p21-dependent signaling pathway in response to g-rays, high level of survivin, increased resistance to cisplatin and UV light and strong expression of the proliferation markers Ki-67 and PCNA (Hawsawi et al, 2008).…”