Vitamin A contents of rat intestinal epithelium and jejunal mucinous adenocarcinoma.

Concentrations of retinol and tetinyl esters were assayed in rat intestinal mucosa and in chemically induced transplanted mucinous adenocarcinoma of the jejunum. Lipid extract from the tissues was chromatographed on deactivated alumina to isolate retinol and retinyl esters, which were determined by specific spectrofluorometry. Normal intestinal mucosa tissue contains 556 ng of retinol equivalents as retinyl esters and 303 ng of free retinol/g of wet tissue. The concentration of retinyl esters in the intestinal mucosa from rats carrying the transplanted tumor was 341 ng/g wet tissue; no free retinol was detected in the small intestinal epithelium of these rats. Liver tissue from the tumor-bearing rats contained 157 microng of retinol equivalents as retinyl esters and 136 microng of free retinol/g of wet tissue. The concentration of vitamin A per cell in the adenocarcinoma tissue was about 20 times less than that in intestinal epithelium.

Expression of breast epithelial differentiation antigens in human primary breast cancer.

Monoclonal antibody LICR-LON-M18 is a marker of normal human breast epithelial cell differentiation. The epitope recognized by LICR-LON-M18 is a prominent component of luminal plasma membranes of nonneoplastic resting and lactating human breast epithelial cells but is rarely expressed by human breast carcinomas. With the use of competitive binding-inhibition studies, the immunodominant portion of the LICR-LON-M18 epitope was shown to be the following oligosaccharide sequence [with galactose (Gal) and N-acetylglucosamine (GlcNAc)]: Gal beta 1----4GlcNAc beta 1----. This structure was distinct from Gal beta 1----3GalNac, which was bound by peanut agglutinin (PNA) and was not recognized by LICR-LON-M18 [corrected]. With the use of biochemical techniques, the present data not only confirmed sialylation and consequent "masking" of the LICR-LON-M18 epitope and PNA determinants in human breast carcinomas but also identified the particular groups of glycoproteins involved in this process. These studies provided additional support for the thesis that sialylation of human breast carcinoma glycoproteins represented an enhancement of specific differentiation events normally regulated in the morphogenesis of nonneoplastic human breast epithelium and that specific glycoproteins became masked during the genesis of primary human breast cancer.

Primary mucinous adenocarcinoma of the endometrium. A clinicopathologic and histochemical study.

Of 256 patients with carcinoma confined to the uterine corpus at the time of hysterectomy treated in the period 1959-1975 at Stanford University Hospital, 98 patients (38%) had neoplasms which demonstrated at least focal intracytoplasmic mucin. In 21 carcinomas (9%), intracytoplasmic mucin production was the dominant form of differentiation--a group which we designate primary mucinous carcinoma of the endometrium. Freedom from relapse and frequency of myometrial invasion were not statistically different for patients whose neoplasms contained intracytoplasmic mucin, regardless of the amount of mucin present, when compared with cases of nonmucin-containing carcinoma. Using histochemical methods, it was impossible reliably to distinguish between the intracytoplasmic mucin produced by carcinomas arising in endometrium and that produced by carcinomas primary in the endocervix. Differential biopsy and fractional curettage are stressed as useful tools in making this clinically important distinction. Since both benign mucinous metaplasia and mucinous carcinoma may arise in the endometrium, it is important to establish histopathologic criteria by which the malignant lesions may be recognized. The use of criteria illustrated in this paper (which include architectural complexity of proliferation, epithelial stratification, loss of epithelial polarity, and nuclear atypicality) resulted in the recognition of mucin producing proliferations which as a group manifest a 50% incidence of myometrial invasion.

Mucinous adenocarcinoma of the prostate.

A case of mucinous adenocarcinoma of the prostate that was diagnosed with the aid of prostate-specific antigen immunoperoxidase staining is reported. Focal areas of the tumor, which were morphologically similar to the remainder of the tumor, stained with neuron-specific enolase by an immunoperoxidase technique and with the Grimelius stain. This tumor is best thought of as a variant of the classic acinotubular adenocarcinoma of the prostate with well-differentiated cells that secrete mucin, rather than as a completely different type of cancer, as proposed previously.

Protooncogene amplification and tumor ploidy in human ovarian neoplasms.

DNA from 24 ovarian tumors, including 16 carcinomas, was examined for amplification of the proto-oncogenes c-myc, int-2, and rc-erbB-2. All cases of carcinoma were also examined by flow cytometry for DNA ploidy and cell cycle analysis, and eight cases of carcinoma were examined for estrogen and progesterone receptors. Protooncogene amplification was not detected in the DNA of benign ovarian neoplasms, or of ovarian carcinomas with low malignant potential. Amplification of c-myc was detected in six of 12 cases of invasive carcinoma, int-2 amplification was present in one case, and c-erbB-2 amplification was not detected in any case. Among the seven cases evidencing protooncogene amplification, three cases showed aneuploidy in tumor DNA, while four showed diploidy. Two cases which showed aneuploidy in tumor DNA did not demonstrate any degree of protooncogene amplification. Protooncogene amplification was frequently associated with morphologic nuclear anaplasia and high mitotic count. Six of the seven cases demonstrating c-myc or int-2 were of the serous type or showed some degree of serous differentiation, while none of the four cases of purely mucinous carcinoma had evidence of amplification. While the total number of cases in the study was limited, it would appear from the trend demonstrated by the data that protooncogene amplification (particularly c-myc) may be involved in the pathogenesis of aggressive common epithelial tumors of the ovary.

S-100 protein positivity in breast carcinomas: a potential pitfall in diagnostic immunohistochemistry.

S-100 protein, originally isolated from neural tissues, has also been identified in various normal and neoplastic cells, including malignant melanomas. A systematic immunohistochemical investigation of this antigen was performed on formalin-fixed paraffin-embedded samples of benign and malignant breast tissues with use of the avidin-biotin-peroxidase complex immunoperoxidase technique and a polyclonal antiserum that recognizes both the alpha and beta subunits of S-100 protein. In benign breast tissue, S-100 protein was present in both epithelial and myoepithelial cells of terminal ducts and lobules; the staining was cytoplasmic and sometimes nuclear. Of 100 randomly selected invasive breast carcinomas, 48 per cent contained S-100 protein-positive tumor cells. Lobular and medullary carcinomas (60 per cent and 80 per cent, respectively) were more frequently positive than ductal carcinomas (45 per cent). Dendritic cells, most likely Langerhans' cells, were present in some carcinomas and were also positive for S-100. There was no relationship of S-100 positivity to histologic differentiation, recurrence interval, or the expression of various tumor markers. The presence of S-100 protein positivity in metastatic breast carcinomas may lead to the erroneous diagnosis of malignant melanoma. Our observations underscore the importance of testing for a broad panel of tumor markers rather than relying on single antigens in evaluating metastatic malignancies of undetermined origin.

Argyrophilic cells in 202 human mucinous breast carcinomas. Relation to histopathologic and clinical factors.

Two hundred two human, mucinous breast carcinomas were investigated for the presence of argyrophilic granules, and these granules were found in 25% of the cases. The granules were located in the cytoplasm and were heterogeneously distributed within the tumors. Tumors with granules were otherwise morphologically indistinguishable from those tumors without granules. The recurrence-free survival was independent of the presence of granules, and no relation was found to other clinical or histopathologic factors. Tumors with granules were found to be estrogen-receptor positive, and they appear to have a slightly less aggressive growth pattern than tumors without granules, but the difference is far from being statistically significant. It is concluded that there is no convincing evidence that this group of primary breast carcinomas with argyrophilia originates from APUD cells.

Oestrogen receptors in mucinous carcinoma of the breast: an immunohistological study using paraffin wax sections.

An immunohistological method (Shintaku-Said method) for the demonstration of oestrogen receptors in routinely processed paraffin wax embedded tissue was applied to 19 cases of mucinous carcinoma of the breast. Seventeen (89%) tumours showed variable degrees of positivity and two were negative. In eight cases the receptors were also assayed biochemically using a dextran-coated charcoal method, and the results of the two methods showed good correlation. No difference in the distribution of positive and negative cases was noted between pure and mixed mucinous tumours, and in the latter group the pattern of staining of the mucinous elements was similar to that seen in the solid elements. It is concluded that the major advantage of this method is its ability to offer for study the distribution of the receptors in individual cells and specific histological structures. The results also indicate that most mucinous carcinomas of the breast are oestrogen receptor positive, irrespective of whether they are pure or mixed type.

Pure and mixed mucinous breast carcinomas: DNA stemline and prognosis.

The DNA stemline of 45 mucinous breast carcinomas was determined by flow cytometry using paraffin embedded archival tissue sections. The material consisted of 26 pure mucinous and 19 mixed mucinous carcinomas. The patients were followed up for at least 15 years or until death. Nearly all pure mucinous carcinomas had a normal DNA stemline (25 of 26) with only one aneuploid tumour. Mixed mucinous carcinomas had a DNA content resembling that of common ductal carcinoma with 11 aneuploid tumours. Aneuploid tumours tended to be of higher grade and stage than diploid tumours. The survival of patients with pure mucinous carcinoma was better than that of patients with mixed mucinous carcinoma. Mucinous carcinoma should be classified as such only if it is a pure mucinous carcinoma.

Influence of mucinous components on survival in colorectal adenocarcinomas: a multivariate analysis.

Of 534 resected colorectal adenocarcinomas, 165 (31%) contained some mucinous components; these represented the main part of the tumour in 67 (13%). Of the mucin containing tumours, 63 (38%) were in the right colon compared with 50 (13%) of the non-mucinous ones (p less than 0.001). Patients with predominantly mucinous tumours were significantly older than those with non-mucinous tumours, and they tended to present with tumours at a more advanced stage. A multivariate analysis did not show any significant independent prognostic influence of the mucinous component except when this had a predominantly signet ring cell pattern.

Distribution of ferritin, transferrin and lactoferrin in breast carcinoma tissue.

An immunoperoxidase staining technique was used for detecting three major iron binding proteins (ferritin, transferrin and lactoferrin) in 40 breast carcinoma cases and six benign breast proliferative lesions. Ferritin staining was detected mainly in connectival stroma and in histiocytes surrounding neoplastic cells. Few and faint ferritin positivities were also detected in neoplastic cells of 20 carcinoma cases. Transferrin was found inconsistently in myoepithelial cells surrounding normal ductules, or around neoplastic ducts of ductal in situ carcinoma. In eight carcinoma cases, transferrin staining was also positive in neoplastic cells. Lactoferrin was detected only in normal breast epithelial cells and in benign breast proliferative lesions. These immunohistochemical findings may suggest that raised serum ferritin concentrations in breast carcinoma patients might be attributed to stromal reaction rather than to tumour synthesis. Transferrin staining of neoplastic cells in these carcinoma cases appears to be very intriguing, particularly since transferrin is considered an obligate requirement for growing cells, and transferrin receptors have been demonstrated only in dividing cells. On the basis of the immunohistochemical data, lactoferrin might be used as a pointer to benign lesions.

S-100 protein in salivary gland tumors: an immunohistochemical study of 129 cases.

Immunohistochemical distribution of S-100 protein was evaluated in 129 tumors from major and minor salivary glands. Also, two sensitive immunoperoxidase avidin-biotin methods using either overnight incubation with primary antibody or pretreatment trypsin digestion and half-hour incubation were compared. Tumors with S-100 protein immunoreactivity were demonstrated in numerous benign and malignant histologic categories. Adenoid cystic carcinomas, carcinomas ex pleomorphic adenoma, clear cell carcinomas, and adenocarcinomas NOS showed inconsistent positive staining, whereas all monomorphic and pleomorphic adenomas and polymorphous low grade adenocarcinomas examined stained positively. No staining was observed in mucoepidermoid carcinomas or acinic cell carcinomas. Mesenchymal-like tumor cells with positive immunostaining were seen only in pleomorphic adenomas and trabecular-tubular adenomas. Equivalent results were found with both overnight and same-day digestion techniques. The consistent S-100 protein staining in some histologic tumor categories (pleomorphic and monomorphic adenoma and polymorphous low grade adenocarcinoma) compared to mucoepidermoid carcinoma that is devoid of S-100 protein immunoreactivity has application to some microscopic differential diagnostic situations. Inconsistent staining of adenoid cystic carcinomas and adenocarcinomas did not allow discrimination from other benign and malignant salivary gland tumors with similar histomorphology.

Estrogen and progesterone receptors in human colorectal tumour cells (study of 70 cases).

Estrogen and progesterone receptors were studied in 70 cases of human colorectal cancer by a cytochemical technique. 28.5% of the cases were estrogen-receptor positive and 42.8% progesterone-receptor positive. There was no difference between the sexes for estrogen receptors but the women had more tumours with progesterone receptors than men. The presence of receptors is unrelated to the differentiation of the tumor. More colon tumours were positive than those of the sigma and rectum. The concentration of cells with receptors in positive cancer cases tended to be low or medium-low.

Correlation of an estrogen receptor-related phosphoprotein with histopathological features in breast cancer.

A series of 65 cases of different histological types of breast carcinoma was investigated for the immunohistochemical location of the estrogen receptor-related, 29 kD phosphoprotein using the ER-D5 monoclonal antibody. The ER-D5 response is heterogeneous in relation to some therapeutic limitations and is correlated with histopathological features of the tumors and survival. The main parameters for evaluation of breast cancers are reviewed, both those that are statistically correlated and those that are not apparently always correlated but are known to have considerable biological meaning, such as the ER-status of tumors.

Immunohistochemical studies on cystic pancreatic neoplasms.

For the treatment of cystic changes of the pancreas, it is essential to distinguish cysts and pseudocysts from neoplasm. Since clinical parameters are usually not characteristic, only histologic and cytologic analyses will prove a diagnosis. The immunohistochemical characteristics of microcystic adenoma, mucinous cystic neoplasia in comparison to solid cystic pancreatic tumor, ductal carcinomas, and endocrine tumors, are studied with a panel of markers as well as enzyme, epithelial, neuroendocrine and pancreatic hormonal markers. The immunohistochemical results with diffuse cytoplasmatic expression of CEA and epithelial markers may be helpful in the exploration of carcinomatous transformed tissue parts within mucinous cystic pancreatic neoplasias, and are therefore of significance for surgical therapy.

Expression of intermediate filaments in different human epithelial and mesenchymal tumors.

Fifty primary gastrointestinal and breast carcinomas, four embryonal rhabdomyosarcomas, six nonmuscular mesenchymal malignant tumors and one mesothelioma have been studied to determine what type of intermediate filaments they express, using affinity purified antibodies to prekeratin, vimentin and desmin and FITC or peroxidase labeled second antibodies. The tissues were alcohol fixed and paraffin embedded before use. In all carcinoma cases the tumor cells are stained by antibodies to prekeratin, while the vimentin antibody only decorates the stroma. Prekeratin positive tumor cells are not only seen in well differentiated tumors, but also in signet ring cell carcinomas. In the case of rhabdomyosarcoma the tumor cells clearly were decorated by antibodies to desmin, while the vimentin antibody only stained very few tumor cells. In cases of nonmuscular mesenchymal tumors, the tumor cells could only be labeled by antibodies to vimentin and not by antibodies to prekeratin or desmin. In biphasic tumors like mesothelioma, different parts of the tumor were separated by antibodies to prekeratin and vimentin.

Prognostic significance of immunohistochemical parameters in breast carcinomas.

136 breast carcinomas were investigated immunohistochemically for alpha-lactalbumin (ALA), lactoferrin (Lfr), human milk fat globule membrane antigen (HMFG-2), transferrin receptor (TrfR), Ki-67 and epidermal growth factor receptor (EGFR). The relationships of pathologic features and steroid receptor status previously shown to be of prognostic significance and the immunohistochemical parameters were examined. It was found that estrogen receptor (ER) was inverse related to TrfR, Ki-67 and EGFR whereas progesterone receptor (PgR) was inverse correlated to Ki-67 solely. Tumor grading revealed significant correlations between TrfR, Ki-67 and HMFG-2 (inverse correlation). Tumor diameter showed correlation between Ki-67 solely. Moreover there were significant relationships between lymphoid infiltration and TrfR, Ki-67 and HMFG-2 (inverse correlation). The comparison of the immunohistochemical parameters showed correlations between Ki-67 and TrfR, Ki-67 and HMFG-2 (inverse correlation) as well as HMFG-2 and ALA. Therefore it is suggested that functional and tumor kinetic properties determined by HMFG-2, Ki-67 and TrfR may be an additional indicator with prognostic significance. On the other hand immunoreactivities of ALA in conjunction with HMFG-2 possibly indicates a subpopulation of breast carcinomas that may have to be investigated further. Moreover these results showed that lymphoid infiltration was correlated with Ki-67 reactivity as well as TrfR expression.

Intermediate-sized filaments in cells of normal human colon mucosa, adenomas and carcinomas.

The distribution of intermediate - sized filaments in human colon mucosa as well as in adenomas and carcinomas of the colon was studied by means of both immunohistology and electron microscopy. The epithelial cells of the colonic mucosa are definitely labelled with antibodies against prekeratin (cytokeratin). Interwoven filaments of the prekeratin type are present in the basal compartments of the epithelial cells; they surround the nuclei and mucus droplets and form an apical skeletal disc. Pericryptal connective tissue is prekeratin negative and vimentin positive. Benign hyperplastic polyps have a high content of prekeratin. The potential precursors of colonic carcinoma, i.e., the tubular and villous adenomas, also show an increase in intermediate-sized filaments of the prekeratin type. Correspondingly, electron microscopy reveals elongated bundles of intermediate-sized filaments arising from the desmosomes of the lateral and basal cell membranes. The prekeratin content is particularly high in adenocarcinomas and highest in mucinous carcinomas. As expected, the stroma of all neoplasms studied is prekeratin-negative, but distinctly vimentin-positive. In one moderately differentiated adenocarcinoma there was evidence of "vimentin-positive" tumor cells. These changes may be caused by binding of cytokeratins with an unknown substance in vimentin antisera, as observed similarly by Moll et al. (1982) in a transitional cloacogenic carcinoma.

The cytokeratin profiles of ovarian common "epithelial" tumors.

An improved immunohistochemical determination of the cytokeratin profiles of epithelia and their neoplasms is possible using monoclonal antibodies that will either identify all 19 cytokeratins (AE1/3) or delineate specific subsets (35 beta H11, 34 beta E12, 34 beta B4 and Cam 5.2). Ovarian common "epithelial" tumors (CET) contain cytokeratin filaments. To determine the nature and differences in the cytokeratin profiles of ovarian CET, eight benign Brenner tumors, four serous cystadenofibromas, 28 mucinous tumors, 27 serous tumors and six endometrioid, five clear cell and five undifferentiated carcinomas, as well as nine normal ovaries were immunostained with the above five antibodies. AE1/3 staining was predominant, while Cam 5.2 and 35 beta H11 displayed the most frequent staining thereafter. Statistically significant staining differences were found between a number of tumor groups using the antibodies 35 beta H11, 34 beta E12 and Cam 5.2. In this study, all ovarian CET, except the benign Brenner tumors, displayed a predominantly low molecular weight cytokeratin profile. The same profile in the normal surface epithelium lends credence to the belief that these tumors are derived from this epithelium. A significant staining difference between some of the tumor types using some of the antibodies suggests a possible ancillary, diagnostic role of cytokeratin profiling in situations where exact tumor typing is difficult.

[Prostaglandin E (PGE) and gastric carcinoma].

Since 1984, an experimental and clinical study on the relation between PGE and gastric carcinoma has been performed by determining PGE content in the bioptic gastric mucosa and plasma. It is found the PGE content in the gastric mucosa and plasma is increased in all patients with gastric cancer, especially with signet ring cell carcinoma. It is higher in the regional lymph node metastasis than in the early cancer, extensive metastases and normal subjects. The PGE content in the plasma is reduced obviously 7-10 days after operation but is increased markedly in recurrent patients. There is no significant difference in extensive metastases, relapse free and normal subjects. The PGE content in the plasma is significantly higher in gastric carcinoma than in chronic atrophic gastritis, but no difference is present between chronic atrophic gastritis and normal subjects.