ABSTRACT
This study sought to understand the role of breast carcinoma-associated fibroblasts in the progression of cancer cells into lymph nodes. We compared fibroblasts of primary tumors and matched the involved lymph nodes to select fibroblast activation markers, namely α-smooth muscle actin (α-SMA), S100A4, and vimentin, as well as to determine the frequency of transforming growth factor ß1, a pleiotropic cytokine that induces the differentiation of fibroblasts to myofibroblasts, and its downstream effectors: CXCR4 and p-AKT. We disposed samples of 80 primary invasive ductal carcinomas and matched the involved lymph nodes from 43 cases into 3 tissue microarrays, and analyzed stromal and tumor epithelial cells separately by immunohistochemistry. Control uninvolved lymph nodes were analyzed by whole-tissue sections. Cancer-associated fibroblast in lymph nodes with macrometastasis expressed similar profiles of vimentin, α-SMA, and S100A4 as those found in primary tumors. Cancer-associated fibroblast were uniformly estrogen receptor, progesterone receptor, HER-2, Ki-67, and p53 negative, but expressions of transforming growth factor ß1 (TGFß1), CXCR4, and p-AKT staining (62.3%, 52.4%, 65%, respectively) were equivalent between primary and lymph node metastasis (LNM) fibroblasts. A significant coexpression of TGFß1 with p-AKT and CXCR4 in LNMs suggested the involvement of these proteins with TGFß1 signaling. These biomarkers, including α-SMA and S100A4, were negative in fibroblasts of cancer-free lymph nodes, with the exception of vimentin. Our finding that expressions of biological markers were similar in fibroblasts of the primary tumors and in matched LNMs, but were absent in cancer-free lymph nodes, supports the assumption that the lymph node stroma mimics the microenvironment observed in primary tumors.
Subject(s)
Biomarkers, Tumor/metabolism , Breast Neoplasms/pathology , Lymphatic Metastasis , Breast Neoplasms/metabolism , Female , Fibroblasts/metabolism , Humans , ImmunohistochemistryABSTRACT
AIMS: Fos-related antigen 1 (Fra-1) is a member of the activator protein 1 (AP-1) transcription factor family. Our objective was to evaluate the role of Fra-1 expression in breast carcinoma progression and prognosis. METHODS AND RESULTS: Fra-1 expression was investigated by immunohistochemistry in two tissue microarrays containing, respectively, 85 ductal carcinoma in situ (DCIS) and 771 invasive ductal carcinoma (IDC) samples. Staining was observed in the nucleus and cytoplasm of the carcinomas, but only nuclear staining was considered to be positive. Fibroblasts associated with IDC were also Fra-1-positive. The frequency of Fra-1 positivity in IDC (22.8%) was lower than that in DCIS (42.2%). No association was found between Fra-1 and clinico-pathological variables in DCIS. In IDC, Fra-1 expression correlated with aggressive phenotype markers, including: high grade, oestrogen receptor negativity and human epidermal growth factor receptor 2 (HER-2) positivity (P=0.001, 0.015 and 0.004, respectively), and marginally with the presence of metastasis (P=0.07). Fra-1 was more frequently positive in basal-like (34%) and in HER-2-positive (38.5%) subtypes than in luminal subtypes. Fra-1 presence did not correlate with survival. CONCLUSIONS: A high frequency of Fra-1 in DCIS tumours may be associated with early events in breast carcinogenesis. Although Fra-1 expression correlated with features of a more aggressive phenotype in IDC, no relationship with overall survival was found.
Subject(s)
Breast Neoplasms/metabolism , Carcinoma, Ductal, Breast/metabolism , Carcinoma, Intraductal, Noninfiltrating/metabolism , Proto-Oncogene Proteins c-fos/metabolism , Adult , Aged , Aged, 80 and over , Antibodies, Monoclonal , Breast Neoplasms/pathology , Carcinoma, Ductal, Breast/pathology , Carcinoma, Intraductal, Noninfiltrating/pathology , Cohort Studies , DNA-Binding Proteins/metabolism , Disease Progression , Female , Follow-Up Studies , Humans , Immunohistochemistry , Middle Aged , Prognosis , Receptor, ErbB-2/metabolism , Retrospective Studies , Tissue Array Analysis , Transcription Factor AP-1/metabolism , Young AdultABSTRACT
Estrogen is essential for growth and development of 30 per cent of malignant breast tumors and concentration of estrogen receptor (ER) is an indicator of hormone dependence. The effects of estradiol-l7beta (E2) on growth promotion and vitamin D (VDR) and progesterone (PR) receptor modulation were evaluated in hormone-dependent (MCF-7, T47D) and -independent (MDA-MB-231) breast carcinoma cells lines. 10(-7)M E2 stimulated the proliferation of MCF-7 and T47D cells the extent of which was correlated to ER content, being sensitive to growth inhibition of 10(-6) M tamoxifen. No effect on growth and PR levels was observed in E2 treated MDA-MB-231 cells. A statistically significant 10(-7)M E(2-)mediated induction of VDR was verified in T47D cells which was abolished by 10(-6)M tamoxifen, revealing an ER-mediated mechanism. 10(-6)M tamoxifen treatment alone upregulated VDR levels in T47D cells, suggesting that this drug may utilize a distinct pathway (ER-independent) for stimulation of VDR content, by a mechanism, now in progress, to be elucidated. These results, taken together, suggest the importance of VDR level assessment as new a prognostic indicator in breast carcinoma.
