Role of HGF in epithelial–stromal cell interactions during progression from benign breast disease to ductal carcinoma in situ

Patricia Casbas-Hernandez,Liza Makowski,Kirk Mcnaughton,Erick Roman-Perez,Jodie M Fleming,Monica D’Arcy,Samantha M Miller,Raghav K Chhetri,Melissa A Troester,Amy L Oldenburg,Heather Ann Brauer,Keith D Amos

doi:10.1186/bcr3476

Abstract

IntroductionBasal-like and luminal breast cancers have distinct stromal–epithelial interactions, which play a role in progression to invasive cancer. However, little is known about how stromal–epithelial interactions evolve in benign and pre-invasive lesions.MethodsTo study epithelial–stromal interactions in basal-like breast cancer progression, we cocultured reduction mammoplasty fibroblasts with the isogenic MCF10 series of cell lines (representing benign/normal, atypical hyperplasia, and ductal carcinoma in situ). We used gene expression microarrays to identify pathways induced by coculture in premalignant cells (MCF10DCIS) compared with normal and benign cells (MCF10A and MCF10AT1). Relevant pathways were then evaluated in vivo for associations with basal-like subtype and were targeted in vitro to evaluate effects on morphogenesis.ResultsOur results show that premalignant MCF10DCIS cells express characteristic gene expression patterns of invasive basal-like microenvironments. Furthermore, while hepatocyte growth factor (HGF) secretion is upregulated (relative to normal, MCF10A levels) when fibroblasts are cocultured with either atypical (MCF10AT1) or premalignant (MCF10DCIS) cells, only MCF10DCIS cells upregulated the HGF receptor MET. In three-dimensional cultures, upregulation of HGF/MET in MCF10DCIS cells induced morphological changes suggestive of invasive potential, and these changes were reversed by antibody-based blocking of HGF signaling. These results are relevant to in vivo progression because high expression of a novel MCF10DCIS-derived HGF signature was correlated with the basal-like subtype, with approximately 86% of basal-like cancers highly expressing the HGF signature, and because high expression of HGF signature was associated with poor survival.ConclusionsCoordinated and complementary changes in HGF/MET expression occur in epithelium and stroma during progression of pre-invasive basal-like lesions. These results suggest that targeting stroma-derived HGF signaling in early carcinogenesis may block progression of basal-like precursor lesions.

Highlights

Basal-like and luminal breast cancers have distinct stromal–epithelial interactions, which play a role in progression to invasive cancer
Cocultures were performed in this media after ascertaining that reduction mammoplasty fibroblasts (RMFs) maintained their RPMI 1640 doubling times in this Dulbecco’s modified Eagle’s medium/F12
Correlation with hepatocyte growth factor (HGF) signature in human tumors We evaluated the behavior of our HGF signature in 707 breast cancer samples from three publically available datasets: NKI295 (N = 295) [18], Naderi and colleagues (N = 135) [19], and UNC337 samples (N = 277) [20]

Summary

Introduction

Basal-like and luminal breast cancers have distinct stromal–epithelial interactions, which play a role in progression to invasive cancer. Basallike breast cancers have a distinct microenvironment interaction pattern relative to other breast cancer subtypes [5] and appear to be associated with distinct immune microenvironments [6,7,8]. These and many other data suggest that complementary epithelial–stromal coevolution is influential in cancer development. Since most of these studies have examined epithelial– stroma interactions after tumors have acquired invasive characteristics, it is not well known how host–tumor interactions are maintained earlier in disease progression

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