Bcl-2 protein in normal, hyperplastic and neoplastic breast tissues. A metabolite of the putative stem-cell subpopulation of the mammary gland.

This investigation, though initially designed to examine the possible influence of the Bcl-2 protein on the node-metastasizing capacity of breast carcinomas, was amplified to study the expression of this anti-apoptotic protein in normal breast lobules and hyperplastic lesions. We examined paraffin sections of 508 breast carcinomas, stained for Bcl-2, estrogen (ER) and progesterone receptors (PgR) and epithelial membrane antigen, and occasionally for other antigens as well. Only a few cells showing a strong Bcl-2 positivity spotted the tubulo-lobular units of normal resting glands, whereas such cells were relatively numerous in atrophic lobules, and very scarce in the terminally differentiated lactating breast. Columnar and usual types of hyperplasia were exclusively, or almost exclusively, composed of Bcl-2(+), ER(+) and PgR(+) cells. The foci of carcinoma in situ and those of invasive carcinomas were respectively 83% and 66% positive for Bcl-2 in at least 25% of their cells. Even among the invasive carcinomas, Bcl-2(+) cases included 83% and 87% of the ER(+) and PgR(+) cases, respectively (p=0.0001). Though there was a statistically significant inverse relation between Bcl-2 and tumor grade (p=0.0001), no significant association was found between Bcl-2 and lymph node stage. In conclusion, we suggest that normal, hyperplastic and neoplastic breast epithelial cells expressing the anti-apoptotic protein Bcl-2 are immature cells that ought to form part of the stem-cell subpopulation, which is committed to the development and to the maintenance of the normal gland and which gives rise to hyperplastic and neoplastic disorders when its proliferation is deregulated. In ductal proliferative changes Bcl-2 assays may be useful for diagnostic but not for prognostic purposes.

EMA: a differentiation antigen related to node metastatic capacity of breast carcinomas.

Epithelial membrane antigen (EMA), also known as MUC1, is a mucinous glycoprotein fixed to the luminal domain of the epithelial cell membrane of normal breast ducts. However, in breast cancer cells, it is usually dispersed in the cytoplasm. EMA staining patterns of 330 breast carcinomas were examined, and three groups formed: lineal (16%), cytoplasmic (75%), and negative (9%). Although these patterns were somewhat related to histological cancer types, this was not statistically significant. However, EMA showed statistically significant univariate relationships to tumor grade, tumor size, estrogen and progesterone receptors, and nodal stage. Logistic regression analysis showed that among these variables, all of which were univariately related to node metastasis, only tumor size and EMA were independent nodal stage predictors. A combined analysis of these two factors revealed that the statistical probability of a tumor metastasizing to four or more nodes increased in each tumor size group from 0.9% to 12% for pT1, from 2% to 29% for pT2 and from 10% to 63% for pT3, depending on the EMA staining. The tumors showing a lineal pattern were the least metastasizing, while the EMA-negative tumors were the most. After recognizing these relationships between EMA staining patterns and other well-known differentiation markers and the lymph node metastatic capacity of carcinomas, and considering the results obtained by others on survival, one might conclude that EMA is both a differentiation marker and a histological prognostic factor.

Importance of estrogen receptors for the behavior of invasive micropapillary carcinoma of the breast. Review of 68 cases with follow-up of 54.

The aggressiveness of invasive micropapillary carcinomas of the breast (MPCa) is still controversially discussed. Therefore, we investigated a total of 68 cases and studied the evolution of 54. MPCa were frequently well-differentiated cancers with the following positivities for immunohistochemistry: 74.5% estrogen receptor (ER+), 46.3% progesterone receptor (PR+), 66% Bcl2+, and 36.4% C-erbB-2+. However, in 90.5% of the cases lymph nodes were involved at diagnosis, and 70.6% of T1 tumors showed wide metastatic spread. After a mean follow-up of 52.6 months, 55.6% of the patients were disease free (DF), 7.4% had disseminated disease and 37% had died. Univariate analysis showed significant differences. Thus, the DF group of patients included 90% of those having tumors with an associated colloid pattern, 73.3% of the ER positive tumors, none of the C-erbB-2 positive tumors, and 100% of the tumors with no axillary metastasis, 77.8% of those with metastasis to up to 3 nodes, and 47.2% of those metastasizing to 4 or more nodes. However, using Cox's regression model for survival analysis, ER was the only factor associated with duration of survival (p = 0.0175). In conclusion, although long-term survival in MPCa is determined by involvement of lymph nodes, as is the case in any other breast cancer type, their short-term evolution is influenced by other factors, mostly by estrogen receptors.

Sialomucins are characteristically O-acylated in poorly differentiated and colloid prostatic adenocarcinomas.

Mucinous glycoproteins are secreted by prostatic adenocarcinomas and might play important roles in tumor invasion and metastasis. Their histochemical properties on routine biopsy specimens have not been fully characterized. We present a histochemical study of mucin in 21 prostatic adenocarcinomas, with particular focus on the demonstration of different types of sialomucins. We applied the following histochemical techniques to routinely processed, formalin-fixed, paraffin-embedded tissue sections: Alcian blue (pH 2.5) and periodic acid-Schiff to reveal both acidic and neutral mucins; high iron diamine and Alcian blue (pH 2.5) to show sulfated and acidic nonsulfated mucosubstances simultaneously; periodic acid borohydride, potassium hydroxide, and periodic acid-Schiff to demonstrate O-acylated sialic acids; periodic acid thionine-Schiff, potassium hydroxide, and periodic acid-Schiff to differentiate pre-existing glycols from those revealed after saponification procedures; and periodic acid borohydride and periodic acid-Schiff to show C9-O-acylated sialic acid. These techniques are useful tools for demonstrating neutral and acidic (sialo- and sulfo-) mucins and di(C8,C9- or C7,C9-)-O-acylated, tri(C7,C8,C9-)-O-acylated and mono(C9)-O-acylated sialomucins. Most prostatic adenocarcinomas showed acidic mucins, with sialomucins predominating over sulfomucins. Well-differentiated and moderately differentiated noncolloid tumors had non-O-acylated sialomucins. Poorly differentiated tumors contained mono-O-acylated (C9) sialomucins, and colloid-type tumors secreted mono-, di-, and tri-O-acylated sialoglycoproteins. Acidic mucins, mainly sialomucins, constitute the major secretory component in prostatic adenocarcinomas, and our results show that the O-acylation of these sialoglycoproteins inversely correlates with tumor differentiation. Well-differentiated and moderately differentiated tumors are not O-acylated, whereas the poorly differentiated ones characteristically have O-acylated sialomucins in C9. Adenocarcinomas of the colloid type, thought to bear a poor prognosis, are the most heavily O-acylated.

Neutral and acid mucins and eosinophil and argyrophil crystalloids in carcinoma and atypical adenomatous hyperplasia of the prostate.

Neutral and acid mucins and eosinophil and argyrophil crystalloids were studied in the glandular intraluminal contents of 130 prostatic carcinomas (PCa) and 22 atypical adenomatous hyperplasias (AAH). Specimens were studied in the usual extension for diagnostic purposes and considered positive when minimal amounts of mucin or any crystalloid were present. Some amount of altered secretion was seen in 110 PCa (84.6%) and in 17 AAH (77.3%); the similarity of the figures speaks for the close relationship of these lesions. The relative incidences in PCa/AAH were 61.5%/77.3% for neutral mucin, 80%/54.5% for acid mucin (p < 0.01), 25.4%/50% for eosinophil crystalloids (p < 0.02), 44.6%/9.1% for argyrophil crystalloids (p < 0.01) and 57%/36% for a compound secretion. The incidence of eosinophil crystalloids was greater in AAH and inversely proportional to the Gleason grade in PCa. The incidence of acid mucins and argyrophil crystalloids was higher in PCa and directly proportional to the tumour grade in all but the poorly differentiated ones. It is therefore suggested that AAH positive for acid mucins or argyrophil crystalloids be classified in the high grade AAH group.

Estrogen and progesterone receptors, c-erbB-2, p53, and Bcl-2 in thirty-three invasive micropapillary breast carcinomas.

Tests for estrogen (ER) and progesterone (PR) receptors, c-erbB-2, p53 and Bcl-2 were made on paraffin sections of thirty-three cases of invasive micropapillary carcinoma (MPCa) of the breast. The relations between these proteins and general parameters and the patients' evolution, were analyzed and their statistical significance determined by Fisher's exact test. Follow up was available on twenty one patients of whom thirteen were alive after a mean of sixty months. Tumor size, metastatic nodes, c-erbB-2 and Bcl-2 all showed higher values in the dead patient group, but only nuclear grade and extensive lymphatic vessel invasion (LVI) were statistically significant prognostic factors. Hormone receptors and oncogenes were positive in quite similar figures to those of common breast cancers (NOSCa) and offered supplementary information about differentiation and cell atypia of individual cancers. Accordingly, ER (72.7%), PR (45.4%) and Bcl-2 (69.6%) were directly interrelated and inversely related with nuclear grade, mitotic grade, c-erbB-2 (36.3%) and p53 (12.1%). In conclusion, MPCa is a lymphotropic cancer phenotype whose prognosis can be influenced by known prognostic factors, including molecular. The lack of discriminative power between MPCa and NOSCa of ER, PR, c-erbB-2, p53, and Bcl-2 reinforces the importance of recognizing this particular type of cancer.

Invasive micropapillary carcinoma of the breast. A new special type of invasive mammary carcinoma.

After reviewing 986 consecutive cases of breast carcinoma from our files, 27 ductal infiltrating cancers showing micropapillary differentiation in invasive areas (MP) were selected. Special immunohistological and ultrastructural techniques were used, and their characteristics compared to those of the Not Otherwise Specified type of carcinomas (NOS). Diagnostic areas of MP were easily identified in H&E sections and were composed of solid or tubular neoplastic cell groups inside a spongy background, where they appeared to swim in aqueous or mucinous material. Neoplastic cells displayed the reverse polarity typical of the papillary phenotype. This was revealed by the detection of acid mucinous rims, lineal deposits of EMA substances, and microvilli in a peripheral position, even in areas where the micropapillae resembled tubules. Histologically, most MP were mixed with N0S, Papillary, or Mucinous patterns, but regardless of the extension of their micropapillary diagnostic component, their tumour size, or their WHO histological grade, two thirds had extensive lymphatic vessel invasion and all the cases presented massive axillary lymph node metastasis. Six of the twelve patients followed died within a mean of 22 months. In conclusion, we propose the recognition of "Invasive Micropapillary Carcinoma of the Breast" as a new special entity with a potentially high degree of aggressiveness.

Argyrophil crystalloids in adenocarcinoma of the prostate.

Considering the advantages provided by the study of glandular secretions in proliferative diseases of the prostate, we carried out a study of 100 consecutive cases of benign hyperplasia (68), glandular dysplasia (12), and carcinoma (20). Samples were predominantly obtained by transurethral resection from patients whose mean ages were 64, 67, and 73.5 years, respectively. The paraffin embedded material was sectioned and stained with hematoxylin-eosin, PAS-diastase, PAS-Alcian blue, methenamine silver, and Masson trichrome. Intraglandular secretion in normal, hyperplastic, and even neoplastic samples was predominantly neutral mucin, but in 2 cases of atypical adenomatous hyperplasia, and in all the carcinomas, we found acid mucin secretion. Holmes crystalloids were present in one case of nodular hyperplasia (1.4%), 2 cases of dysplasia (16.6%), and in 5 carcinomas (25%). As another manifestation of altered secretion, we also saw numerous needle-shaped argyrophilic and isotropic crystalloids, not previously described, in 12 of the 20 carcinomas (60%). These crystalloids were exclusively found in the carcinomas, reached a size of 50 mu, and were completely differentiated from the Holmes crystalloids by their odd needleshape and their staining characteristics.

Carcinoma arising in a calcified bronchogenic cyst.

A round calcified mass located in the anterior mediastinal compartment was removed in a 42-year-old man. The histopathological diagnosis was bronchogenic cyst with foci of anaplastic carcinoma and areas of osseous metaplasia. The rarity of these findings and the possibility of malignancy in bronchogenic cysts is discussed.