[Immunohistochemical study of cancer-associated carbohydrate antigens in carcinoma of the biliary tract].

To reveal the cell-biological character of biliary tract cancer, localization and distribution of three cancer-associated carbohydrate antigens (CA19-9, sialyl SSEA-1, NCC-ST-439) and carcinoembryonic antigen (CEA) were studied immunohistochemically in 35 cases of gallbladder carcinoma, 21 of bile duct carcinoma, 16 of chronic cholecystitis, and 3 of normal gallbladder. 1) All carbohydrate antigens and CEA were present in 70-90% of the cases of gallbladder and bile duct carcinoma. In particular, NCC-ST-439 had the highest incidence of positive staining (95.2%) in bile duct carcinoma. 2) The mode of localization was diverse and was not fixed by the kind of antigen. Antigens flowing out to the surrounding stroma were affected by the rate of positive cells. 3) No significant correlation was observed between the histological type or degree of differentiation and tissue positivity. 4) The positivity of tissue CEA was higher in the cases with serous membrane invasion, gamma INF pattern, and neuro-, vascular-, and lymphatic invasion. 5) In chronic cholecystitis, CA19-9, NCC-ST-439, and CEA were stained in mucosal cells and/or metaplastic cells, while sialyl SSEA-1 was stained only in one case in the goblet cells and the cells with pseudopyloric metaplasia. None of the antigens were stained in normal gallbladders. These results suggest that these antigens may be useful in the diagnosis and therapeutic treatments in patients with biliary tract cancer.

Pathophysiology of tumor progression in human gallbladder: flow cytometry, CEA, and CA 19-9 levels in bile and serum in different stages of gallbladder disease.

Gallbladder epithelium is unique among the gastrointestinal cell types because proteins and protein levels in the fluid bathing the luminal side of the cells (bile) are different from and can be compared with those in the fluid bathing the basal side (serum). To help identify cellular changes that occur during the development of gallbladder cancer, we obtained gallbladder tissue, serum, and bile specimens from 20 patients with invasive adenocarcinoma of the gallbladder, three with high-grade dysplasia (carcinoma in situ), six with low-grade dysplasia, 12 with hyperplasia, and 10 with acute or chronic cholecystitis. We obtained serum samples from 40 patients with invasive adenocarcinoma and bile samples from 29 of these patients; serum samples from three with high-grade dysplasia and bile specimens from two of these; serum and bile samples from five with low-grade dysplasia; serum or bile samples from 126 with metaplasia, hyperplasia, or cholecystitis, including serum samples from 121 and bile samples from 110; and serum and bile samples from eight with normal biliary tracts. The study was conducted in Mexico City, Mexico, and La Paz, Bolivia. We performed flow cytometric DNA analysis on gallbladder tissue specimens and measured levels of carcinoembryonic antigen (CEA) and CA 19-9 antigen in the serum and bile specimens. Analysis of the cell cycle compartments by flow cytometry revealed marked variations of the proliferation index for the different disease states (P less than .0001). The proliferation index increased with progression from cholecystitis to invasive adenocarcinoma. Of the bile and serum measurements, only serum CA 19-9 values were correlated with flow cytometry measurements (r = -.49, P = .005). Overall, the serum and bile measurements were in agreement (P less than .01). However, with the exception of the correlations among serum measurements for the patients with invasive adenocarcinoma, most of the correlations could be explained by differences in the disease state. In particular, the progression from normal tissue to invasive adenocarcinoma involved no change in bile CA 19-9 level and only a slight change in bile CEA level but much larger changes in serum CEA and CA 19-9 levels. It appears that the progression from normal tissue to invasive adenocarcinoma results in increased production of these antigens and often in loss of cell polarity as well, i.e., inability to prevent leakage of the antigens into the serum.

Carcinoid tumors of the gallbladder with adenocarcinomatous differentiation: a morphologic and immunohistochemical study.

Two patients with gallbladder carcinoid tumors with adenocarcinomatous differentiation were examined. In both cases, the tumor contained argyrophilic granules and alcian blue-positive mucin. One contained argentaffin granules and the other showed PAS-positive mucin. Numerous membrane-bound electron-dense neurosecretory granules were demonstrated by ultrastructural study. Immunohistochemistry applied for the tumors clarified the epithelial, hormonal, and metaplastic character. Epithelial tumor markers, i.e., carcinoembryonic antigen (CEA) and epithelial membrane antigen (EMA), were positive in these tumors. The neuroendocrine nature was demonstrated by positive results of chromogranin A and neuron-specific enolase (NSE). Hormonal activities were not confirmed in the tumor cells. These results suggested that carcinoid tumors in the gallbladder have a multidirectional differentiation represented by a morphological continuum ranging from carcinoid to adenocarcinoma.

Estrogen and progesterone receptors in gallbladder cancer.

Cancerous tissues from 21 patients with primary gallbladder cancer were examined immuno-histochemically for the presence of receptors for estrogen (ER) and progesterone (PGR). ER and PGR, localized in the nucleus, were evident in 52.4 per cent and 0 per cent of the patients, respectively. Furthermore, ER and PGR were positive only in the cytoplasm of cancer cells in 28.6 per cent and 66.7 per cent, respectively. There was a higher tendency of moderately- and poorly-differentiated adenocarcinoma to have an ER-positive rate than well-differentiated adenocarcinoma. With respect to the relationship between ER and sex, ER-positive nuclei were observed in 8 of 14 women (57.1 per cent) and 3 of 7 men (42.9 per cent), but the difference between the two was not significant due to the small number of subjects. These result suggested that gallbladder cancers with ER in the nuclei may respond to antihormone therapy.

Epithelial markers in synovial sarcoma. An immunohistochemical study on paraffin embedded tissues.

Immunohistochemical studies on synovial sarcomas have proved the potentiality of these neoplasm for epithelial and mesenchymal differentiation and antibodies detecting epithelial cells have been found to be helpful in determining the histological types. In this study different epithelial markers directed against various cytokeratins, HMFG-2 and EMA were investigated on paraffin embedded tissues of 13 cases of synovial sarcomas, with regard to their reliability in unmasking the epithelial components demonstrable in this type of neoplasm. The results lead to three conclusions: firstly, synovial sarcomas possess the capacity for generating different epithelial cell types with uncommon compositions of intermediate filaments as well as of membrane proteins, secondly, these features may be expressed in a heterogenous pattern even within the same tumour and finally, the use of wide range anti-cytokeratin antibodies covering the spectrum of basic as well as acidic type proteins seems to be necessary for the detection of all epithelial components demonstrable in synovial sarcomas.

Plasminogen activators and plasminogen activator inhibitor in malignant and non-malignant ascitic fluid.

Ascitic fluid from tumour patients (hepatoma, gastric cancer, gallbladder cancer, colorectal cancer, ovarian cancer) and from non-malignant diseases (liver cirrhosis, congestive heart failure) were compared with respect to their content of determinants of the fibrinolytic system, tissue-type plasminogen activator antigen (t-PAag) and activity (t-PAact), urokinase-type plasminogen activator antigen (u-PA) and plasminogen activator inhibitor activity (PAI). Furthermore, SDS-polyacrylamide slab-gel electrophoresis (SDS-PAGE) was performed to evaluate molecular weight distribution of the detectable fibrinolytic parameters. In malignant ascites, PAI activity was three to four times higher, and increased complex formation of PAI with t-PA could be demonstrated, compared with non-malignant ascitic fluid. Tissue-type plasminogen activator antigen and activity showed a similar concentration in ascites of both study groups. Urokinase-type plasminogen activator antigen was detectable neither in ascites of malignant nor in ascites of non-malignant origin. It is concluded that t-PA is the physiological plasminogen activator in ascites and that increased PAI levels followed by increased complex formation between t-PA and PAI might reflect a reaction of the peritoneum.

[Nuclear DNA pattern and various prognostic factors of the gallbladder cancer].

Nuclear DNA patterns which was obtained by the Feulgen staining and microspectrophotometric measurement in the gallbladder cancer were analyzed. DNA score was determined by the peak ploidy unit and decided numbers of frequency in each ploidy unit and compared with the various factors influencing prognosis of the patients. DNA score was significantly lower in cancer with papillary form and limited involvement to the mucosa compared with that with other forms and involvement beyond the proper muscle layer. DNA score was also significantly lower in cancer without hepatic infiltration, vascular invasion and lymph node metastasis than that with those characteristics. In experimental studies VX2 cancer cells which were implanted in the gallbladder subserosa in rabbits, DAN score did not show any changes with the growth of carcinoma implanted. Biologic nature of the tumor which was evaluated with nuclear DNA pattern of cancer cells was fundamental to define the spread of carcinoma and the prognosis of the patients.

[Estrogen receptors in human gastric, hepatocellular, and gallbladder carcinomas and normal liver tissues].

Steroid binding assay using the dextran coated charcoal (DCC) method was applied to human tissues including tumors of the digestive organs, and the results were compared with those of enzymeimmunoassay (EIA) and immunocytochemical assay (ICA) with monoclonal antibody against human estrogen receptor of MCF-7 breast cancer cells. Using the DCC method, estrogen receptor activity was detected in 6 of 26 cases (23.1%) with gastric carcinoma, 3 of 16 hepatocellular carcinoma cases (18.8%), 1 of 3 gallbladder carcinoma cases (33.3%), and both of the 2 cases (100%) with normal liver tissue. However, using EIA, no ER activity was detected in any case. Moreover, ER positive cells were not found by immunohistochemical staining in the gastric carcinoma cases or in normal liver tissue, both of which showed ER activity by the DCC method. These results suggest that the estrogen receptor like material exists in cytosol of the human digestive tumors and normal liver tissue, but that the specificity of the antibodies against estrogen receptor molecules in these tumors may be different from that of the breast tumors.

Undifferentiated carcinoma of the gallbladder. A clinicopathologic, histochemical, and immunohistochemical study of 21 patients with a poor prognosis.

Among 284 cases of carcinoma of the gallbladder, 21 were identified as undifferentiated carcinoma (UC), with little glandular or other specific epithelial differentiation. These tumors were classified into three histologic types according to the components: (1) small cell type (eight cases); (2) pleomorphic cell type (eight cases); and (3) spindle cell or pseudosarcomatous type (five cases). Histochemical and immunohistochemical study by the immunoperoxidase technique revealed that most of the tumors (13/21) contained mucosubstances, and that all examples of the UC were immunoreactive for epithelial membrane antigen (EMA), keratin, and carcinoembryonic antigen (CEA), thereby indicating the epithelial nature of the neoplastic cells. Vimentin immunoreactivity was found in nine tumors. In 19, the tumor contained various neoplastic endocrine cells, including somatostatin-immunoreactive (14/19), gastrin-immunoreactive (14/19), human chorionic gonadotropin (HCG)-immunoreactive (9/19), pancreatic polypeptide-immunoreactive (4/19), and serotonin-immunoreactive cells (4/19). The prognosis of patients with UC of the gallbladder was poorer than that of patients with differentiated adenocarcinoma.

Adenosquamous carcinoma of the gallbladder with spindle cell features. A light microscopic and immunocytochemical study of a case.

A case of adenosquamous carcinoma of the gallbladder showing extensive spindle transformation is presented. By light microscopy, areas showing interwoven fascicles of fusiform, poorly differentiated cells closely resembling a sarcoma were seen to merge imperceptibly with areas showing more obvious glandular and squamous cell features. Immunocytochemistry utilizing tissue-specific antibodies against intermediate filaments demonstrated the exclusive presence of prekeratin antibodies in both components of the tumour, thus establishing the epithelial nature of this neoplasm. The importance of immunological phenotyping in the differential diagnosis of epithelial tumours of the gallbladder showing pseudosarcomatous features is underscored.

Cáncer de la vesícula biliar: estudio anatomoclínico de 100 casos / Gallblader cancer: anatomoclinical study of 100 cases

Se estudian retrospectivamente 100 casos de cáncer vesicular operados en el Hospital Clínico de la Pontificia Universidad Católica de Chile entre 1970 y 1984. Se registra un aparente aumento de su incidencia en los últimos años. Es mas frecuente en el sexo femenino, sobre los 50 años y se asocia a colelitiasis en el 87% de los casos. Más del 85% son adenocarcinomas. Su diagnóstico con frecuencia se hace por laparotomía y corresponde a etapas irresecables, en las que sólo caben medidas paliativas. Su pronóstico, generalmente es muy malo con sobrevida a 5 años menor al 5%. Los pocos casos con mejor sobrevida corresponden a pacientes con "cáncer oculto", de tipo papilar y en los que el tumor no sobrepasa la serosa. Se propone como única medida práctica actual para mejorar su pronóstico, la detección y tratamiento intraoperatorio del "cáncer oculto". Se recomienda la colecistectomía profiláctica en pacientes de mayor riesgo

[Is vesicular cholesterolosis a particular anatomo-clinical form of cholesterol lithiasis? Apropos of a case of cholesterol polyp manifested by an acute complication]. / La cholestérolose vésiculaire est-elle une forme anatomo-clinique particulière de la lithiase cholestérolique? A propos d'une observation de polype cholestérolique révélé par une complication aiguë.

A case of cholesterolic polyp, revealed by the formation of an infundibulo-cystic enclave, is reported, the polyp having the appearance of a cholesterolic calculus, but a pediculated one, and one acting as a calculus. The extreme rarity of this complication is emphasized. Despite the frequency of associated lithiasis, the cholesterolosis are included in a different framework from lithiasis, that of the cholecystoses that on histology show typical pure parietal lesions without inflammation. Findings in this atypical case suggest possible common pathogenic factors for the two affections, and raise a proposal for inclusion of cholesterolosis within the nosologic framework of lithiasis.

Colony-stimulating factor producing carcinoma of the gallbladder.

A patient with an anaplastic carcinoma of the gallbladder accompanied by marked neutrophilia was found to have high colony-stimulating factor (CSF) activity in the urine and serum. The supernatant of the cultured tumor cells (6th passage) showed high and specific CSF activity. These specimens induced the formation of granulocyte colonies, macrophage colonies or granulocyte-macrophage mixed colonies by both human and C57BL mouse bone marrow cells in soft agar. These results suggest that this tumor is a human granulocyte-macrophage (GM)-CSF producing tumor.

The localization of human keratin proteins at cytological and histological levels in carcinomatous and sarcomatous lesions.

Immunoperoxidase staining for human keratin proteins was performed cytologically on samples from 90 patients with malignant tumors, and histologically on samples from 164 patients with malignant tumors. At the cytological level, almost all tumor cells not only in squamous cell carcinoma but also in nonsquamous cell carcinoma were positive for keratin proteins, in contrast with the apparent abscence of keratin proteins in sarcoma. At the histological level, almost all neoplastic cells of squamous cell carcinoma were positive for keratin proteins, the same as at the cytological level. In contrast, among cases of nonsquamous cell carcinoma, the frequency of appearance of keratin proteins varied according to the organ; it tended to be low in tumors with relatively good prognosis, such as carcinomas in the digestive system or thyroid cancer, and to be high in tumor with poor prognosis, such as pulmonary cancer, gallbladder cancer and endometrial cancer. However, there was a marked difference between the frequency of appearance of keratin proteins at the cytological level and that at the histological level, particularly in the cases of gastric cancer.

Immune status in advanced upper gastrointestinal cancers.

Thirty-eight cases of advanced upper gastrointestinal cancer were assessed for their immune status prior to any form of therapy. Cell-mediated immunity, as tested by absolute lymphocyte count, T-cell count and delayed cutaneous hypersensitivity reactions to purified protein derivative and phytohemagglutinin showed severe depression. Immunoglobulins A and M were elevated, while immunoglobulin G reduced. No correlation could be established between the immune status and the site of cancer or its extent, the performance status of the patients, or their response to chemotherapy.

Identification of immunoreactive thyrotropin-releasing hormone in human neoplasia.

Immunoreactive TRH (IR-TRH) was identified in extracts derived from seven human cancer tissues, including two oat cell carcinomas. The IR-TRH concentrations ranged from 29-189 pg/g. Tumor IR-TRH exhibited immunological identity with synthetic pyroglu-his-proamide and coeluted with synthetic [3H]TRH after high pressure liquid chromatography on a mu Bondapak C-18 column. These data extend the number of hypothalamic peptide hormones identified in human neoplastic tissues.

An immunocytochemical study of the distribution of lysozyme, a1-antitrypsin and a1-antichymotrypsin in the normal and pathological gall bladder.

We have studied the distribution of lysozyme (Ly), a1-antitrypsin (a1AT) and a1-antichymotrypsin ( a1AChy ) in the normal, chronically inflamed and neoplastic gall bladder mucosa using the peroxidase-anti-peroxidase (PAP) method. Ly was absent from the normal mucosa but it was found only in areas of glandular metaplasia of true antral type and in crypts of possible early metaplastic nature in cases of chronic cholecystitis. a1AT and a1AChy were also found in such metaplastic areas, but their presence was also observed immunohistochemically in areas of essentially normal and in non-metaplastic, chronically inflamed gall bladder mucosa. The possible local production of these substances by gall bladder epithelial cells is discussed. Ly, a1AT and a1AChy were also found in various histological types of adenocarcinoma of the gall bladder in varying degrees of frequency and intensity, unrelated to the histological type and invasiveness of the tumour.

Alpha-1-antitrypsin (AAT) deposits in gall bladder adenocarcinoma and liver in partial AAT deficiency (Pi SZ phenotype).

Alpha-1-antitrypsin (AAT) immunoreactive inclusions were found in an adenocarcinoma of a gall bladder of a patient with Pi SZ phenotype who also had globular AAT accumulation in the liver. The inclusions did not react with antibodies against other plasma proteins, thus suggesting a primary synthesis of AAT in these cells. The presence of AAT in the tumor cells may represent resurgence of an oncofetal antigen, or, indirectly, may mean that AAT-containing cells normally exist in the human non-neoplastic gall bladder. The accumulation of AAT in tumor cells might be a basic process in malignant transformation.

Sulfatide cholecystosis.

Polypoid masses metachromatic sulfatides have been found at autopsy in the gallbladder of patients with metachromatic leucodystrophy. In a 10-year-old girl with this disease oral cholecystrography demonstrated a filling defect, which was felt to represent a polyp. In the proper clinical setting, sulfatide cholecystosis should be considered in the differential diagnosis of polypoid lesions of the gallbladder.

Polysaccharides of metaplastic mucosa and carcinoma of the gallbladder.

The polysaccharide composition of the human gallbladder well was studied in carcinomas and metaplastic changes of various degrees, and the results obtained were compared with those for the normal material previously presented (Terho, T., and Laitio, M. Biochim. Biophys. Acta 338: 135, 1974). Elevated amounts of acid connective tissue polysaccharides (heparitin and dermatan sulfates as well as chondroitin 4- or 6-sulfate, or both, could be observed in carinomas. In histochemical stainings it was found that in carcinomas and in the two specimens classified as group III (containing the most extensive metaplastic changes at disposal), the intracellular mucin was mainly neutral or nonsulfated acidic. The amounts of sulfated mucin were relatively insignificant. This mucin polysaccharide material was isolated and its composition was determined. It was observed to be large polysaccharide material was isolated and its composition was determined. It was observed to be large molecular (approximate molecular sizes 1 to 2 times 10-6), and to be composed of fucose, galactose, glucosamine, and galactosamine as well as small amounts of sialic acid. The basic structure of these polysaccharides is thus similar to that of normal sulfated mucin. The almost total absence of acid groups, however, causes the polysaccharide material in question to stain in a manner identical with neutral mucin when investigated with histochemical methods. The carcinomas also contained some sulfomucin; its proportion, however, was small as compared with the amounts of nonsulfated acid and neutral mucin in biochemical characterization. A small molecular polysaccharide fraction, assumed to originate in membrane-bound glycoproteins, was isolated from the insoluble gallbladder tissue residue. The proportion of this fraction was larger in carcinomas than in normal material. This rise as well as the rise in the quantity of acid connective tissue polysaccharides is presumably due to the large number of cells in the carcinoma tissue as well as to fibrosis.