Correlation of carbonic anhydrase IX expression with clinico-morphological parameters, hormonal receptor status and HER-2 expression in breast cancer.

UNLABELLED: The hypoxia-inducible protein carbonic anhydrase IX is widely expressed in most human cancers, including breast carcinomas. CA IX attracts significant interest due to its strong association with neoplasms and its absence from corresponding normal tissues, suggesting its potential to serve as a promising diagnostic biomarker. This protein comes into the limelight also as a valuable prognostic and predictive parameter. Immunohistochemically, we examined the expression of this protein in 84 cases of invasive breast carcinoma to determinate the association with clinico-morphological and biological parameters such as age of patients, grade, stage and size of primary tumor, lymph node metastasis, vascular invasion as well as hormone receptor status and HER-2 expression. In each case, the subcellular localization of CA IX antigen, the intensity of staining and the percentage of labeled cells were assessed. Overall, CA IX was expressed in 34 cases (40.5%). The statistical analysis revealed a significant correlation between subcellular localization of CA IX and the age of patients. Furthermore, significant correlations were also found between the grade, estrogen and progesterone status and all immunohistochemical characteristics of CA IX expression (the subcellular localization of CA IX antigen, the intensity of staining and the percentage of labeled cells). We point out that mostly membrane or combined membrane and cytoplasmic positivity together with a higher intensity of CA IX immunoreactivity are associated with poor prognostic parameters, such as tumor grade 3 and also with negative estrogen and progesterone receptor status which may influence therapeutic approach. However, no significant correlations were shown with remaining clinico-morphological and biological parameters. We next investigate the relationship between CA IX expression in the group of invasive ductal carcinomas and the group of invasive lobular carcinomas and other less frequent types of breast carcinoma. There was, however, no significant difference. Our results suggest that moderate to strong membrane and combined membrane and cytoplasmic localization of CA IX may represent a valuable tumor biomarker as well as a promising prognostic and predictive parameter in invasive breast cancer. KEYWORDS: breast carcinoma, carbonic anhydrase IX, immunohistochemistry, clinico-morphological and biological parameters.

The normal human newborns thymus.

The thymic microenvironment constitutes a unique cell environment composed of thymic epithelial cells, myoid cells, and bone marrow-derived accessory cells for the differentiation, maturation and selection of T lymphocytes. The histological feature of thymus is markedly dependent on the age of individual and on various negative stimuli. Our study group consisted of fourteen newborns whose thymuses were removed during surgery performed for various congenital heart defects. We used a palette of seven monoclonal antibodies for exact localization of different cells creating the thymic microenvironment (cytokeratin AE1/AE3, desmin, actin, S100 protein, CD68, CD20, and CD45RO) as well as three monoclonal antibodies against proteins regulating the process of apoptosis (bcl2 oncoprotein, p53 protein, and survivin). We described and microphotographically illustrated the localization of thymic cytokeratin AE1/AE3-positive epithelial cells (subcapsular part of the cortex and medulla, especially Hassall's corpuscles), dendritic cells (medulla, often inside the Hassall's corpuscles), thymic myoid cells (medulla, often in close contact with Hassall's corpuscles), macrophages (mostly cortex, but also medulla and inside the Hassall's corpuscles), B lymphocytes (thymic medulla) and CD45RO-positive T lymphocytes (mostly thymic cortex). We found p53-positive thymic epithelial cell nuclei in subcapsular part of cortex and in outer epithelial cell layer of Hassall's corpuscles (very similar to the basal layer of epidermis). Bcl2 positive lymphocytes were mostly localized in thymic medulla, especially nearby Hassall's corpuscles. The thymuses were mostly survivin-negative with exception of round cells in border between cortex and connective tissue septa (probably migrating progenitor cells) (Tab. 1, Fig. 14, Ref. 66).

The thymus: picture review of human thymus prenatal development.

The thymus is derived from pharyngeal region; a region from which, in case of aquatic vertebrates, the gills develop. According to the classical hypothesis, the epithelial thymus stroma of human embryos is derived from the endodermal cells of the left and right ventral parts of the third pharyngeal pouches. But a close contact of the third pharyngeal pouch with its corresponding third pharyngeal clefts ectoderm plays an important role. Also an epithelial-mesenchymal interaction between the third pharyngeal pouch endoderm and surrounding neural crest derived-cells ectomesenchyme is necessary for the proliferation and differentiation of thymic epithelial cells. In our work we photographically presented the development of thymus from the 6th up to the 25th week of development. The first primordia of the thymus and parathyroid glands within the endoderm of third pharyngeal pouches can be seen in 8 to 9 mm stages. We found also an epithelial proliferation in the second pharyngeal pouches, but this "thymus secundus" stopped their differentiation. The thymus primordia at the 7th and 8th week of development contain almost exclusively epithelial cells. These cells are arranged at the periphery as a row of prismatic cells. The mesenchyme accumulates around the epithelial thymic primordium, and during 9th to 12th weeks of development, septa from mesenchyme fold between the epithelial cells and create the "openings" in the capsular surround. According to our observations, in the 13th week of development the differentiation of cortex and medulla becomes obvious and is completed from the 17th up to 18th week of development onward. The first developing Hassall's corpuscle was detected in the 13th week of development. The striking increase in the number of the Hassall's bodies was observed between the 16th and the 18th week of development, as well as between the 22nd and the 25th week of development (Fig. 14, Ref. 35).