Gene expression profiling of tumour epithelial and stromal compartments during breast cancer progression.
ABSTRACT The progression of ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) marks a critical step in the evolution of breast cancer. There is some evidence to suggest that dynamic interactions between the neoplastic cells and the tumour microenvironment play an important role. Using the whole-genome cDNA-mediated annealing, selection, extension and ligation assay (WG-DASL, Illumina), we performed gene expression profiling on 87 formalin-fixed paraffin-embedded (FFPE) samples from 17 patients consisting of matched IDC, DCIS and three types of stroma: IDC-S (<3 mm from IDC), DCIS-S (<3 mm from DCIS) and breast cancer associated-normal stroma (BC-NS; >10 mm from IDC or DCIS). Differential gene expression analysis was validated by quantitative real time-PCR, immunohistochemistry and immunofluorescence. The expression of several genes was down-regulated in stroma from cancer patients relative to normal stroma from reduction mammoplasties. In contrast, neoplastic epithelium underwent more gene expression changes during progression, including down regulation of SFRP1. In particular, we observed that molecules related to extracellular matrix (ECM) remodelling (e.g. COL11A1, COL5A2 and MMP13) were differentially expressed between DCIS and IDC. COL11A1 was overexpressed in IDC relative to DCIS and was expressed by both the epithelial and stromal compartments but was enriched in invading neoplastic epithelial cells. The contributions of both the epithelial and stromal compartments to the clinically important scenario of progression from DCIS to IDC. Gene expression profiles, we identified differential expression of genes related to ECM remodelling, and specifically the elevated expression of genes such as COL11A1, COL5A2 and MMP13 in epithelial cells of IDC. We propose that these expression changes could be involved in facilitating the transition from in situ disease to invasive cancer and may thus mark a critical point in disease development.
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ABSTRACT: Prognostication is an integral part of cancer diagnostic and helps oncologists to guide treatment decisions and therapy intensity. Accumulating evidence suggest that the stroma compartment also contains independent prognostic information, best exemplified by the impact of immune cells and cells of the vasculature on cancer progression. Similarly, strong experimental evidence exist that stromal fibroblasts, often designated cancer associated fibroblasts (CAFs), are actively involved in tumorigenesis. Thus, it can be anticipated that the molecular repertoire of CAFs is likewise important for the clinical behaviour of the tumor. In this review we present recent studies addressing the prognostic impact of CAFs, with the focus on human lung and breast cancer. Several single markers have been suggested, either CAF specific or CAF derived, that in immunohistochemical studies have demonstrated independent association with survival. This includes members of the platelet derived growth factor receptor (PDGFR) family, CAF-markers like podoplanin and fibroblast activation protein (FAP) as well as transcription factors (FoxF1) and secreted factors (matrix metalloproteinases (MMPs), SPARC). However, most studies are based on explorative evaluations on single patient cohorts and require further validation. Using a more comprehensive approach, microarray studies have been employed to create gene expression signatures that detect an activated fibroblast state. These “stroma signatures have been applied to identify specific CAF features associated with prognosis in several independent data sets of breast and lung cancer patients. Early studies in breast cancer have also demonstrated that fibroblast features influence therapy response. Thus, many strategies have been used to present encouraging proof-of-concept findings that CAFs could be exploited for prognostication. However, these studies also highlight the difficulties to conclusively define an “activated stroma and to identify the individual factors involved in clinically relevant tumor-stroma interactions.Seminars in Cancer Biology 01/2014; · 7.44 Impact Factor
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ABSTRACT: Vav1 is one of the signalling proteins normally restricted to hematopoietic cells that results ectopically expressed in solid tumors, including breast cancer. By immunohistochemical analysis on TMAs containing invasive breast tumor from patients without lymph node involvement, we have found that Vav1 is expressed in almost all investigated cancers and shows a peculiar localization inside the nucleus of tumor cells. High amounts of nuclear Vav1 are positively correlated with low incidence of relapse, regardless phenotype and molecular subtype of breast neoplasia. In particular, Kaplan-Meier plots showed an elevated risk of distant metastasis in patients with low Vav1 expression compared with patients with high Vav1 expression in their tumors. Experiments performed with breast tumor-derived cells indicated that Vav1 negatively modulates their invasiveness in vitro and their metastatic efficiency in vivo, possibly by affecting the expression of genes involved in invasion and/or metastasis of breast tumors. Since the high heterogeneity of breast tumors makes difficult to predict the evolution of early breast neoplasias, the evaluation of nuclear Vav1 levels may help in the characterization and management of early breast cancer patients. In particular, Vav1 may serve as a prognostic biomarker and a target for new therapies aimed to prevent breast cancer progression.Oncotarget 05/2014; · 6.64 Impact Factor
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ABSTRACT: CCN (Cysteine-rich protein 61, Connective tissue growth factor and Nephroblastoma overexpressed gene) comprise a family of matricellular proteins that have multiple physiologic functions including development, tissue repair, cell adhesion, migration and proliferation. The expression of CCN1, cyclin D1, beta-catenin and p53 was explored by immunohistochemistry in different grades of Ductal Carcinoma in Situ (DCIS) cases. These cases did not contain any infiltrating carcinoma components. In addition, all CYR61 gene exons (encoding the CCN1 protein) were sequenced in 30 samples. Allred and H-scores were calculated for expression in both DCIS and the surrounding benign breast tissue. All cases of DCIS showed degrees of cytoplasmic CCN1 staining with median H-scores of 170, 160 and 60 in grades 3, 2 and 1 respectively (p = 0.043). Twelve of 28 DCIS 3, 1 of 15 DCIS 2 and 0 of 18 DCIS 1 also showed nuclear staining for CCN1. The cytoplasmic staining difference was preserved when the cases were divided into ER+/DCIS grade 1, ER+/DCIS2&3 and ER-/DCIS2&3 by the H-score (p = 0.037). Cyclin D1 expression was positively correlated with the CCN1 cytoplasmic H-score in all DCIS samples (p = 0.038). Membranous beta-catenin expression correlated with the grade of intraepithelial carcinoma by both H-score (p = 0.047) and Allred Score (p = 0.026). Our results suggest that CCN1 has a role in development of intraepithelial carcinoma. CCN1 expression correlates with grade of DCIS independent of ER status. It can induce cell-cycle progression through cyclinD1. It is warranted to study high expression of CCN1 in DCIS as an independent risk factor in a larger cohort.Human pathology 01/2014; · 3.03 Impact Factor