FAS is a reliable prognostic marker to predict DFS and OS in patients with early breast cancer.

Purpose: hMena (ENAH), a cytoskeleton regulatory protein involved in the regulation of cell motility and adhesion, is overexpressed in breast cancer. The aim of this study was to define at what stage of breast carcinogenesis hMena is overexpressed and to correlate hMena overexpression with established prognostic factors in breast cancer, focusing on human epidermal growth factor receptor-2 (HER-2). Experimental Design: hMena expression was assessed immunohistochemically in a prospective cohort of cases (n = 360) encompassing a highly representative spectrum of benign breast diseases associated with different risk of transformation, in situ, invasive, and metastatic tumors. Correlations with conventional pathologic and prognostic variables, such as proliferation index, hormonal receptor status, and HER-2 overexpression, were also evaluated. In vitro experiments were done to study the effect of neuregulin-1and Herceptin treatments on hMena expression. Results: hMena protein is undetectable in normal breast and is weakly expressed in a small percentage of low-risk benign diseases (9%), but displays a progressive and significant increase of positivity in benign lesions at higher risk of transformation (slightly increased risk 43%; moderate increased risk 67%), in in situ (72%), invasive (93%), and metastatic breast cancer (91%).A significant direct correlation with tumor size (P = 0.04), proliferation index (P < 0.0001), and HER-2 overexpression (P < 0.0001) and an inverse relationship with estrogen (P = 0.036) and progesterone receptors (P = 0.001) are found in invasive carcinomas. In vitro experiments show that neuregulin-1up-regulates, whereas Herceptin down-regulates, hMena expression. Conclusions: Our data provide new insights into the relevance of actin-binding proteins in human breast carcinogenesis and indicate hMena overexpression as a surrogate indicator in breast disease management. Management of breast cancer, and other malignancies, islikely to benefit from the identification of early markers of transformation. In this context, major efforts are ongoing in the areas of genomic and proteomic profiling aimed at identifying genetic or biochemical markers related to proliferative lesions heralding the development of breast cancer. In view of the increasing evidence that the host immune response contributes to the editing of the tumor phenotype (1), we have recently isolated, by serologic analysis of cDNA expression libraries (SEREX), hMena (ENAH) protein, the human orthologue of murine Mena, which is overexpressed in over 70% of primary breast cancers (2). Mena belongs to the Ena/VASP protein family, which, by controlling the geometry of the actin filament network (3, 4), represents key regulator molecules of cell movement and shape in a large variety of cell types and organisms (5). It has been suggested that Ena/VASP proteins are members of the adherens junction structures and are required for the actin dynamics necessary to seal membranes into epithelial sheets (6), a process frequently deregula...

Breast cancer is a heterogeneous disease, and within the HER-2 positive subtype this is highly exemplified by the presence of substantial phenotypical and clinical heterogeneity, mostly related to hormonal receptor (HR) expression. It is well known how HER-2 positivity is commonly associated with a more aggressive tumor phenotype and decreased overall survival and, moreover, with a reduced benefit from endocrine treatment. Preclinical studies corroborate the role played by functional crosstalks between HER-2 and estrogen receptor (ER) signaling in endocrine resistance and, more recently, the activation of ER signaling is emerging as a possible mechanism of resistance to HER-2 blocking agents. Indeed, HER-2 positive breast cancer heterogeneity has been suggested to underlie the variability of response not only to endocrine treatments, but also to HER-2 blocking agents. Among HER-2 positive tumors, HR status probably defines two distinct subtypes, with dissimilar clinical behavior and different sensitivity to anticancer agents. The triple positive subtype, namely, ER/PgR/Her-2 positive tumors, could be considered the subset which most closely resembles the HER-2 negative/HR positive tumors, with substantial differences in biology and clinical outcome. We argue on whether in this subgroup the "standard" treatment may be considered, in selected cases, i.e., small tumors, low tumor burden, high expression of both hormonal receptors, an overtreatment. This article review the existing literature on biologic and clinical data concerning the HER-2/ER/PgR positive tumors, in an attempt to better define the HER-2 subtypes and to optimize the use of HER-2 targeted agents, chemotherapy and endocrine treatments in the various subsets.

Screening of a cDNA expression library from a primary breast tumor with the autologous patient serum led to the isolation of 6 cDNA clones corresponding to 3 different genes, including a novel gene that maps to chromosome 1 and encodes the human homologue of mouse Mena (hMena, cDNA clone RMNY-BR-55), a protein of the Ena/VASP family involved in the regulation of cell motility and adhesion. A cancer-restricted antibody response against hMena was demonstrated, since 18/93 cancer patient sera, the majority (10/ 52) from breast cancer, showed anti-hMena-specific IgG, while no antibodies were present in healthy donors. When hMena protein expression was analyzed by Western blot and immunohistochemistry, the antigen was overexpressed in the majority of breast cancer cell lines and in 75% of primary breast tumor lesions evaluated. Furthermore, when HLA-A2-restricted peptides from the hMena sequence were used to stimulate CD8 ؉ T cells, an hMena-specific response was found in 9 out of 12 HLA-A2 ؉ breast cancer patients. In 4 patients, this cell-mediated immune response was concomitant with antibody response to hMena. Furthermore, an hMena-specific T-cell line was established from an HLA-A2 Key words: tumor immunity; major histocompatibility complex; cytotoxic T lymphocyteThe identification of the repertoire of molecules recognized by the immune system of cancer patients at different stages of the disease is of major biologic and clinical relevance, 1 as the immune system continuously shapes the immunogenic phenotype of the developing tumor 2 by a complex process recently referred to as cancer immunoediting. 3 Genetic changes continuously occurring during tumor development and progression lead to a number of mutant and/or aberrantly expressed proteins, which can potentially function as tumor-associated antigens and elicit antitumor immune responses. 4 However, the dynamics and the consequences of these events have not yet been fully elucidated. The serologic analysis of cDNA expression libraries (SEREX) of human tumors has identified a broad spectrum of tumor proteins capable of eliciting a humoral immune response in tumor patients. 5 The majority of these SEREX-defined antigens do not show any detectable mutations and/or structural modifications. Although some tumor antigens show restricted expression in normal tissues, i.e., cancer/testis (CT) and melanoma differentiation antigens, to date, the results indicate that the overexpression of normal proteins in the tumor may be of major significance in eliciting a tumor-specific humoral immunity. 6 The isolation of tumor antigens recognized by high-titer IgG implies CD4 ϩ and CD8 ϩ T-cell recognition, as extensively demonstrated for the C/T antigen NY-ESO-1 that induces a concomitant humoral and cellular immune response in a high proportion of NY-ESO-1 ϩ cancer-bearing patients. [7][8][9][10] To identify and characterize new antigens in breast cancer, we applied the SEREX approach analyzing a primary breast tumor and the autologous serum of a long surviving patient. A novel...

Tenascin (TN) is a high-molecular-mass oligomeric glycoprotein of the extracellular matrix (ECM) endowed with a programmed expression in embryonic life and a neo-expression in the interstitium of some malignancies. Using monoclonal antibodies (MAbs) which identify human tenascin, we have conducted an extensive immunohistochemical analysis of TN expression in normal fetal and adult human tissues as well as in a wide variety of human tumors. Results of this study demonstrate that TN (1) is detectable in embryonic and fetal tissues at least from the 10th week of gestation; (2) is present in the interstitium of a variety of adult tissues of different embryonic origin; (3) may be neo-expressed in the stroma of benign and malignant tumors; (4) has the ability to accumulate in a highly variable manner in the ECM of tumors of the same and of different histotypes.