Melanoma-associated antigen (MAGE) family members are generally described as tumor-specific antigens. An association between MAGE-D4B and breast cancer has yet to be reported and the functional role of the encoded protein has never been established. We performed microarray analysis of 104 invasive breast tumors and matched non-cancerous breast biopsies. qPCR was used for validation in an independent biobank. To investigate the biological relevance of MAGE-D4B in breast tumorigenesis, its phenotypic effects were assessed in vitro. Overall, MAGE-D4B was detected in 43% of tumors while undetected in normal breast tissue. MAGE-D4B was found to correlate with tumor progression and to be an independent prognostic marker for poor outcome in term of relapse-free and overall survival, with potential predictive relevance in relation to response to chemotherapy. RNA interference-mediated knockdown of MAGE-D4B significantly hampered the invasive properties of Hs578T cells by affecting anchorage-independent growth, adhesion, migration and invasion affecting anchorageindependent growth, adhesion, migration and invasion and by modulating expression of invasion-suppressor gene E-cadherin.Breast cancer is the most common cancer in women and one of the leading causes of death 1 and it is characterized by high heterogeneity at the histopathologic and molecular levels, which is ultimately reflected in the clinical course of the disease and responses to treatment. The prognosis and clinical management of patients with breast cancer are commonly determined by clinicopathological factors and immunochemical markers. Histological type, grade, tumor size, lymph node involvement, estrogen (ER) and progesterone (PR) receptors status and HER2-receptor overexpression/gene amplification all influence prognosis and probability of response to systemic therapies; however, they fail to fully capture the varied clinical course of breast cancer. 2 Expression of human melanoma-associated antigen (MAGE) family genes has been recently associated with several types of cancer. [3][4][5][6][7] The first human MAGE family member was discovered as a gene encoding a tumor-specific antigen. 8 Since then, more than 50 MAGE genes have been identified and classified as type I and type II genes, based on differences in gene structure and tissue-specific gene expression. 9 Type II MAGE-D genes are characterized by a more complex genomic structure compared to type I genes, with an open reading frame encoded by multiple exons. 10 One of these genes, MAGE-D4B, has been reported in human malignancies, including gliomas, non-small-cell lung cancers and oral squamous cell carcinomas. [11][12][13] However, an association of MAGE-D4B expression with breast cancer has not previously been established and the functional role of the encoded protein remains uncharacterized.In this study, by genome-wide microarray analysis of a large cohort of patients, we identified MAGE-D4B to be differentially expressed in invasive breast tumors compared with normal breast tissue and to be ...

Context.-Loss of 1 copy of chromosome 3 is considered a significant indicator of metastatic dissemination in uveal melanoma. Fresh or paraffin-embedded tumor tissue is most commonly used for current cytogenetic techniques for determining chromosome 3 status in uveal melanoma and often requires referral to an external specialist laboratory for analysis.Objectives.-To assess the chromogenic in situ hybridization assay for detecting chromosome 3 alterations using frozen tumor imprints and to compare the results obtained with those obtained by standard fluorescence in situ hybridization or single-nucleotide polymorphism array techniques.Design.-Chromogenic in situ hybridization was performed on 52 frozen uveal melanoma tumor imprints. The genetic status of 26 of the 52 cases had been determined previously by fluorescence in situ hybridization (group 1); the status of 26 cases had been determined using singlenucleotide polymorphism array (group 2).Results.-Chromogenic in situ hybridization was successfully performed on 48 of 52 tumor imprints. Chromogenic in situ hybridization showed excellent agreement in all 24 cases determined by fluorescence in situ hybridization (100% concordance; j ¼ 1; P , .001; 95% confidence interval, 100%-100%), and disagreed in 4 of the 24 cases previously studied by single-nucleotide polymorphism array (83% concordance; j ¼ 0.67; P , .001; 95% confidence interval, 95%-39%). All 4 discordant cases were classified as disomic for chromosome 3 by chromogenic in situ hybridization and monosomic by SNP array. On histologic examination, the 4 discordant cases corresponded to 2 mixed cell tumors and 2 spindle cell tumors.Conclusions.-Chromogenic in situ hybridization using tumor imprints is a reliable technique for determining chromosome 3 status in uveal melanoma. Furthermore, it can also be easily integrated into a routine histopathology laboratory.