Imprint immunocytochemistry of estrogen receptors in breast carcinoma.

To clarify the accuracy of immunocytochemical detection of estrogen receptors (ER) in breast carcinomas using cytological materials, imprint specimens from tumor tissue were compared with frozen tissue sections and tumors analyzed by the dextran coated charcoal (DCC) method and enzyme immunoassay (EIA). Out of 50 cases examined by imprint immunocytochemistry, there were 39 ER positive cases (78.0% positivity). The positivity in the imprint materials agreed with that of the DCC in 36 out of 40 cases (85.0%), with 100% sensitivity and 60.0% specificity. The two methods statistically correlated with each other in their positivity and grade (p less than 0.001). The positivity and grades of imprint and frozen immunocytochemistry as well as those of imprint immunocytochemistry and the EIA agreed almost perfectly with each other. As a result of the present study, we concluded that immunocytochemical detection of ER is indeed reliable, as accurate as other procedures. We recommend that aspiration biopsy cytology (ABC) be used for morphological examination and ER immunocytochemistry when adequate materials are available and that imprint materials be used when ABC materials are inadequate and fresh tissue is available at the time of surgery.

Two homologous mixed müllerian tumor lines of the ovary and their characteristics.

Two cell lines, NF and JoN, derived from human ovarian carcinosarcomas, were established in tissue culture and in nude mice. Both lines, growing in monolayers, showed morphologic features of adenocarcinoma cells (NF being aneuploid with a modal number of 53, and JoN being pseudodiploid with a modal number of 44). Intermediate filaments were demonstrated immunohistochemically; the JoN line expressed keratin, but not vimentin or desmin, whereas the NF line expressed vimentin and desmin, but not keratin. Plasminogen activator activity was found in both lines. It is concluded that both of these lines are potentially useful models for studying the diverse characteristics of malignant mixed Müllerian tumors.

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.

Regional ploidy variations in signet ring cell carcinomas of the stomach.

Regional ploidy variations within individual tumors were analyzed by in-situ cytofluorometry of metaphase cells in Feulgen-stained paraffin sections, using 45 resected stomachs with early and advanced signet ring cell carcinomas. Aneuploid cells were found in one of 30 early cancers and in eight of 15 advanced cancers, and were almost always accompanied by diploid cancer cells in the mucosal part of the cancers. The diploid and the aneuploid cells were generally found to be distributed in different territories in the mucosa, and aneuploid foci were often included in the diploid area. These findings suggest the diploid origin of signet ring cell carcinomas and the occurrence of aneuploidy during the tumor development. Moreover, the aneuploid cells appeared to infiltrate beyond the mucosa more readily than the diploid cells; most of the aneuploid populations already invaded the extramucosal tissue, and the cancer cells infiltrating in the extramucosal tissue were predominantly aneuploid in six of the nine cancers with aneuploidy. Thus, it appears that the occurrence of aneuploid clones may accelerate the progression of signet ring cell carcinomas from early to advanced stages.

Correlation between tumor histology, steroid receptor status, and adenosine deaminase complexing protein immunoreactivity in ovarian cancer.

Adenosine deaminase complexing protein (ADCP) immunoreactivity was investigated in 40 ovarian tumors and correlated with clinicopathologic parameters, including tumor steroid receptor content. Ten (29%) of 34 common epithelial ovarian carcinomas showed ADCP reactivity. Reactivity for ADCP was seen more frequently in mucinous (100%; p less than 0.001), well-differentiated (73%; p less than 0.001) and Stage I (56%; p less than 0.05) ovarian carcinomas. Furthermore, tumors that contained high levels of androgen receptors and tumors that did not contain estrogen receptors were more frequently ADCP positive (p less than 0.05). However, after stratifying for histologic grade, no correlation between ADCP reactivity and receptor status was found. Determination of ADCP reactivity appears to be of limited value in ovarian cancer.

[A clinicopathological study of signet-ring cell carcinomas of the stomach].

Surgically resected signet-ring cell carcinomas of the stomach have been clinicopathologically investigated. Although this type of carcinoma was found to be widely spread in the propria mucosa, a deeper invasion beyond the submucosa appeared more slowly than in other types of carcinomas. For example, the larger the early carcinoma in which the invasion was restricted to within the submucosa, the greater the incidence of a signet-ring cell carcinoma increase, especially in cases involving the mucosa. The incidence of a nodal metastasis was found to be lower than in cases of a moderately and poorly-differentiated adenocarcinoma in the early stage. However, when the signet-ring cell carcinomas invaded beyond the submucosa, the tumor cells spread rapidly and widely in the wall with a subsequent abdominal implantation, causing ascites and peritonitis carcinomatosa. It seemed likely that this deep invasion was accelerated by cellular change, such as the enlargement of nucleus, cellular atypy, and a decreased mucin production. As signet-ring cell carcinoma arise from the neck of glands and infiltrate the propria mucosae under the superficial epithelium, diagnosis by the barium enema and an endoscopic examination is very difficult in the case of small-size lesions. In lesions greater than 1 cm in diameter, these carcinomas usually showed an erosive and/or ulcerated appearance.

[A study of localizations and serum data; four tumor markers (CA125, CA19-9, CEA and TPA) in ovarian cancers].

Four tumor markers (CA125, CA19-9, CEA, TPA) were analysed between the localizations and the serum data in the 36 ovarian cancers. The positive rates of each tumor markers were; 23 cases (63.9%) on CA125, 15 cases (41.7%) on CA19-9, 12 cases (33.3%) on CEA, and 27 cases (75.0%) on TPA. CA125 and CA19-9 were observed in the luminar borders and cellular membranes on serous adenocarcinomas and in the cytoplasms on mucinous adenocarcinomas. Both CEA and TPA were stained in cytoplasms. The correlation in the localization was observed between CA19-9s and CEAs (Kendall's rank correlation = 0.5303 greater than 0.5). The correlations between the localization and the serum data were observed on CA19-9 and CA125.

Expression of ras oncogene p21 protein in early gastric carcinoma and adjacent gastric epithelia.

The expression of the ras gene product p21 in normal gastric mucosa, early gastric carcinoma of diffuse (gastric) and intestinal types, and in adjacent mucosal abnormalities is reported. The analysis was performed on paraffin sections by an immunohistochemical assay using the mouse monoclonal antibody RAP-5 and the rat monoclonal antibody Y13-259. Expression of ras p21 was assessed by staining intensity and percentage of positively stained cells. In comparison to normal gastric mucosa of non-cancer patients, p21 was overexpressed in nearly all early carcinomas of both types and in the dysplastic and/or metaplastic mucosal alterations accompanying intestinal type of gastric cancer. Increased p21 expression was also observed in the normal-appearing mucosa adjacent to early carcinomas of diffuse type, but not in the morphologically normal gastric epithelium adjacent to the intestinal type. The results of this investigation suggest that ras p21 overexpression may be related to early events of human gastric carcinogenesis. The study supports the notion of different pathways in the development of diffuse (gastric) and intestinal types of gastric carcinomas.

Ovarian mucinous tumor with mural nodules of anaplastic carcinoma.

A case of mucinous cystic ovarian tumor with mural nodules of anaplastic carcinoma in a 30-year-old woman is described. The carcinomatous components within the nodules showed strong immunopositivity for cytokeratin and carcinoembryonic antigen, and ultrastructurally they displayed epithelial and glandular differentiation. Omental metastasis had already developed in the patient, and she received postoperative adjuvant chemotherapy consisting of cyclophosphamide and cis-platinum. No sign of recurrence was evident 4 months after the operation. The literature is reviewed and the importance of adjuvant chemotherapy in the postoperative management of such patients highlighted. The salient pathologic features differentiating mural nodules of anaplastic carcinoma and true sarcoma from prognostically favorable sarcoma-like nodules are presented.

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.

[A clinicopathological study of early mucinous adenocarcinoma of the stomach].

From 1963 to 1986, 1126 cases (1300 lesions) of early gastric carcinoma were treated at Fukui Prefectural Hospital. Mucinous adenocarcinomas comprised 16 lesions in this series. These cases of mucinous adenocarcinoma were younger than all the other early carcinomas of the stomach and they often were locted in the lower third of the stomach. Macroscopically, most were types "Iia + IIc". Dividing them into 3 grouping: papillary, tubular, and signet ring cell types, depending on the microscopic appearance of the intramucosal carcinoma, the signet ring cell group differed from the other two as to age, macroscopic appearance, and region it occupied. An immunochemical study showed that CA19-9 stained positively in a mucous lake, much like a carcinoma cell, but that CEA stained little.

[Relation of the serosal invasion mode to a recurrence in advanced gastric carcinomas].

Discussed are the attributable factors affecting the type of carcinomatous recurrence seen in 126 patients who had been operated on for their gastric carcinomas from 1979 to 1982 and who later showed a macroscopically positive serosal invasion (S0) and a histologically ss alpha approximately se invasion and required a curative resection. Forty-six percent of the patients with an ss beta approximately se invasion had a peritoneal recurrence, 21% a liver recurrence, and 33% a recurrence of some other type, whereas those with ss alpha had no such recurrence. Peritoneal recurrence tended to increase with the increase in the length of the serosal invasion and its rates were: one third in serosal invasions of less than 3 cm, one third in invasions of 3 approximately 6 cm and another third in invasions of over 6 cm. The smaller ratio of submucosal length to subserosal length, especially when less than 1.0, meant a greater frequency of peritoneal recurrence. The histologic characteristics of carcinomas that developed a peritoneal recurrence were poorly differentiated, contained INF alpha and had weaker cellular cohesion, whereas those that developed a liver metastasis were well differentiated carcinomas, contained INF beta and had a tighter cellular cohesion.

[Signet-ring cell carcinoma of the urinary bladder: study of 4 cases and review of the literature]. / Carcinoma de células en anillo de sello de la vejiga urinaria: estudio de cuatro casos y revisión de la literatura.

Four cases of primary signet-ring cell adenocarcinoma of the urinary bladder are described. All patients were men, with ages ranging from 46 to 81 years. The number of signet-ring cells varied and showed a direct correlation with poor prognosis. PAP analyses of acid phosphatase and prostate-specific antigen were negative. The results show that signet-ring cell carcinoma has a poor prognosis. This cell type should be sought in all bladder carcinomas since its presence is likely to worsen prognosis.

[A case of mucinous carcinoma of the skin].

We were consulted by an 81-year-old man who had been complaining of a slow-growing tumor on his abdomen for 20 years. Histologically, small islets of the tumor cells were floating in mucinous lake separated by fibrous septa, so we diagnosed this tumor as mucinous carcinoma of the skin (Mendoza). Electron-microscopically, nuclei of the tumor cells were slightly folded. There were a few secretory granules and many secretory vacuoles in cytoplasm, but no findings of decapitation secretion. Like some other authors' reports, our data suggest that this tumor has an eccrine gland origin. To our knowledge, this is the 12th case of mucinous carcinoma of the skin in the Japanese literature.

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.

Carcinoma of the pancreas: a clinicopathologic study of 96 cases with immunohistochemical observations.

We carried out clinicopathological and immunohistochemical studies on tissues from 96 patients with pancreatic carcinoma, excluding cystic neoplasms (cystadenocarcinomas). The cumulative three-year survival rate was 8.8%, and 58 patients (60.4%) died within one year of the initial surgery. The survival curves for patients with pancreatic carcinoma with no venous invasion and no lymph node metastasis were significantly more favorable than for those with venous invasion and/or lymph node metastasis. The curves for patients with well-differentiated adenocarcinoma were significantly more favorable than for those with moderately or poorly differentiated adenocarcinoma. Fifteen patients with small pancreatic carcinomas, the greatest diameter being less than 3 cm, had more favorable postsurgical outcomes, with a cumulative two-year survival rate of 45%, compared to 81 patients with more advanced carcinomas, with a rate of 11% (P less than 0.05). In the majority of cases, the tumor contained carcinoembryonic antigen, carbohydrate antigen 19-9 and Ha-ras p21 immunoreactants in cell membranes or cytoplasm of the cells and/or in the stroma. There was no significant relationship between the immunohistochemical results and the clinical outcome.

Distribution of fibronectin and laminin in early and advanced signet-ring-cell carcinomas of the stomach.

An immunoperoxidase method was used to compare the distribution of fibronectin and laminin between superficially spreading and deeply infiltrating parts of signet-ring-cell carcinoma of the stomach. In both parts, laminin-containing basement membranes were generally scarce, but they were observed on some of the cancer cells which had differentiated to glandular cells. Intramucosal invasion fronts of superficially spreading cancers often showed a layered structure, consisting of a middle zone of small cancer cells together with a superficial and a deep zone of signet-ring cells. In this structure, linear fibronectin and laminin deposits were common on the cancer cells in the deep zone, but rare in the superficial zone. However, fibrillar fibronectin deposits in the stroma were not considerably larger in either zone. At extramucosal invasion fronts of deeply infiltrating (advanced) cancers, a stromal remodelling with an increased amount of fibrillar fibronectin deposits was often observed around the cancer cells, whose cell surface fibronectin was largely lost even from some basement membranes. These findings suggest that invasive activity of signet-ring-cell carcinoma may not be related to the mere presence or absence of cell-surface fibronectin and laminin but to the amount of stromal fibronectin, which could reflect a cell-stroma interaction. Signet-ring-cell carcinomas have a stage of intramucosal growth in which cancer cells may live in dependence upon the pre-existing stroma and form the layered structure, while in advanced stages cancer cells seemed to have acquired an ability to elicit their own "tumor stroma".

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.

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.