Biological characterization of pseudocyst-forming cell lines from human adenoid cystic carcinomas of minor salivary gland origin.

Two cell lines (ACCS and ACCY) were isolated from two individuals with adenoid cystic carcinoma (AdCC) using tissue culture techniques. Both cell lines have similar morphology, i.e., elongated and flattened cells with slender cytoplasmic processes. The two cell lines tend to form pseudocysts, which are a specific architectural feature of AdCC. Coexpression of cytokeratin and vimentin was found in the two cell lines, which occasionally also contained S-100 protein and lactoferrin or lysozyme immunoreactivity. Moreover, ACCS and ACCY displayed potential for the production of a large amount of extracellular matrix including basal lamina components such as fibronectin, laminin, and type IV collagen and glycosaminoglycans which are also part of the basal lamina. These findings suggest that the tumor cells, probably basal or myoepithelial like cells, are responsible for the formation of the peculiar stroma of AdCC consisting of a large amount of collagen-like fibers, basal lamina components, and mucopolysaccharides.

Basement membrane laminin and type IV collagen in various benign and malignant adnexal tumors of the skin: an immunohistochemical study.

Thirty benign and seven malignant adnexal tumors of the skin and one lymph node metastasis were stained for laminin and type IV collagen with rabbit antibodies against the human basement membrane (BM) proteins using the immunoperoxidase technique. Fifteen of the benign sweat gland, sebaceous gland, and hair follicle tumors showed a continuous and distinct BM around the tumor aggregates. The cylindromas and eccrine spiradenomas seemed to produce excessive amounts of BM material, part of which was seen as amorphic patches within the tumor cell clusters, whereas the trichofolliculomas, trichoepitheliomas, and pilomatrixomas showed an absence of BM from many areas. In syringomas, in addition to the tubular structures surrounded by a continuous BM, undifferentiated cell nests containing granular BM material were present. They probably represent primitive structures obtaining during early development into tubules. The seven malignant tumors and the only metastasis studied here all contained small, narrow strips of BM material extracellularly between the infiltrating tumor clusters. Only in two cases was faint staining for laminin found within the cells. The pepsin pretreatment of the formalin-fixed, paraffin-embedded samples had most probably degraded the intracytoplasmic BM material in most cases. The BM defects were found to be associated with malignancy and low differentiation of the adnexal skin tumors, as reported previously for other tumor types, but a partial loss of BM was also associated with high differentiation in some benign adnexal tumors.

"Adenoid cystic" carcinoma and adenoid basal carcinoma of the uterine cervix. A study of 28 cases.

Adenoid cystic carcinoma (ACC) and adenoid basal carcinoma (ABC) of the uterine cervix are rare tumors that have often been regarded as a single entity. We studied 28 cases of these neoplasms, with 14 cases in each category. Most patients were over 60 years of age, and there was a high proportion of black women. The majority of the women with ACC presented with postmenopausal bleeding and had an obvious mass on pelvic examination. Despite the tumors' architectural similarity to ACC of the salivary gland, microscopic examination of the cervical carcinomas showed necrosis, a high mitotic rate, and greater nuclear pleomorphism. In all but one of the cases, the tumor cells were negative for S-100 protein on immunoperoxidase staining--a finding that provides evidence against a myoepithelial component. However, S-100-positive dendritic cells were present in the stroma of the tumors and among the neoplastic cells. The patients with ABC were usually asymptomatic, without a gross abnormality of the cervix. Microscopic examination disclosed small nests of basaloid cells, almost always beneath, and often arising from, in situ or small invasive squamous cell carcinomas. In contrast to ABC, ACC was often complicated by local recurrence or distant metastasis. We conclude that ACC of the uterine cervix differs from ACC of salivary gland origin and is also distinct clinically and pathologically from cervical ABC.

Adenoid cystic carcinoma of the trachea and main-stem bronchus. A clinical, histopathologic, and immunohistochemical study.

Twelve cases of adenoid cystic carcinoma of the trachea and main-stem bronchus were histologically analyzed, and the results were examined with reference to the growth pattern of the tumor and the prognosis. The tumors were histologically classified into tubular, cribriform, and solid subtypes. Three histologic grades were established: grade I, tumors with tubular and cribriform subtypes but without solid subtype; grade II, tumors with tubular and cribriform subtypes in which the solid subtype comprised less than 20% of the area; grade III, tumors in which the solid subtype comprised more than 20% of the area. Three gross infiltrating types were established: type I, entirely intraluminal; type II, predominantly intraluminal; type III, predominantly extraluminal. In most cases histologic grade correlated with gross tumor type; that is, grades, I, II, and III were grossly types I, II, and III, respectively. The tumors infiltrating along the tracheobronchial wall were of the tubular or cribriform subtype, but not of the solid subtype. In two patients who died of distant metastasis, the histologic studies revealed the solid subtype. Immunohistochemical analysis demonstrated that the tubular subtype was the most differentiated form and the solid subtype, the most undifferentiated form. The histologic subtype of adenoid cystic carcinoma of the tracheobronchial tree was an important factor in the growth pattern of the tumor and the prognosis.

Adenoid cystic carcinoma of the uterine cervix: ultrastructure, immunofluorescence, and criteria for diagnosis.

An example of adenoid cystic carcinoma of the cervix was recently encountered in our laboratory and studied by histochemistry, electron microscopy, and immunofluorescence in order to compare the neoplasm with adenoid cystic tumors at other sites and to establish criteria for diagnosis. Histochemically, cervical adenoid cystic carcinoma showed the two types of mucin, epithelial and stromal, as expected in adenoid cystic carcinomas of other organs. Ultrastructurally, this tumor was characterized by redundant basal lamina forming pseudocysts, intercellular spaces, and occasional true lumens with microvilli. Immunofluorescence studies showed that the cells contain at least two antigenically different types of filaments, actin and keratin, and that the cells produce true basement membrane (collagen IV). The presence of actin suggests myoepithelial differentiation even though the tumor probably originated from the cervical reserve cells, and myoepithelium is not a known component of normal cervix. This study shows that cervical adenoid cystic carcinoma is a distinct entity which can be identified and separated from other types of cervical adenocarcinomas.

Adenoid cystic carcinoma. A clinicopathologic study with flow cytometric analysis.

Forty-five patients with adenoid cystic carcinoma (ACC) of salivary glands were retrospectively studied to determine the prognostic use of DNA ploidy analysis compared with clinicopathologic features. Nuclear suspensions were prepared from 37 of these tumors by the Hedley technique on paraffin-embedded material. The DNA content was analyzed by flow cytometry after propidium iodide staining. Thirty-five tumors were diploid and 2 were tetraploid. Mean survival was 117 and 52 months for the diploid and tetraploid patients, respectively. The median S-phase fraction (Spf) of the 35 diploid tumors was 4.45%. Of the 17 diploid patients who died of disease, there were 11 tumors with high Spf (greater than 4.45%) and 6 tumors with a low Spf (P less than 0.05 chi-square test). The presence of more than 30% of a solid pattern in the tumor was strongly associated with poor median survival in Kaplan-Meier survival curve analysis (P less than 0.05 log rank test). Grade, stage, recurrence, and metastases were also found to be important prognostic factors. Because few tumors were nondiploid, these results suggest that S-phase fraction analysis may be a more useful prognostic indicator than ploidy classification.

Isolation and sequence analysis of human bombesin-like peptides.

The decapeptide form of human gastrin releasing peptide was isolated from acid extracts of liver tissue containing a metastatic human bronchial carcinoid tumor. A larger form also was isolated and partially characterized. During gel permeation chromatography the major immunoreactive peak eluted in the same region as synthetic gastrin releasing decapeptide while a second minor immunoreactive peak eluted near gastrin releasing peptide. Bombesin-like immunoreactivity (BLI) was purified by successive applications to reverse phase high pressure liquid chromatography (HPLC) columns. After four successive HPLC purifications a single peak of bombesin-like immunoreactivity was detected. Amino acid analysis, microsequence analysis and coelution with synthetic peptide indicated that the predominant form present in metastatic tumor tissue was identical to the decapeptide form of canine gastrin-releasing peptide. The less abundant form was purified by cation exchange chromatography followed by reverse phase high pressure liquid chromatography. Partial microsequence analysis of this peptide, through the first 11 residues, was Val-Pro-Leu-Pro-Ala-Gly-Gly-Gly-Thr-Val-Leu. This sequence differed from that of hog heptacosapeptide gastrin releasing peptide at positions 1,3,4 and 5 and from the canine peptide as positions 1,3,5, and 7.

Antibodies to intermediate filament proteins. The differential diagnosis of cutaneous tumors.

One hundred cutaneous tumors were investigated immunohistopathologically for the expression of intermediate filament (IF) proteins. Epithelial tumors, such as basocellular and squamous cell carcinomas, cutaneous adnexal tumors, and metastatic carcinomas showed keratin positivity in a varying number of tumor cells with two keratin antibodies with different specificities. Neoplastic cells of fibrohistiocytic tumors, pigmented nevi, melanomas, hemangiomas, glomus tumors, and lymphomas were positive for vimentin, but not for keratin or desmin. Cutaneous leiomyomas and leiomyosarcomas, on the other hand, were positive for desmin. The results show that the typing of IFs enables the differential diagnosis between carcinomas and sarcomas or melanomas, epidermal appendage tumors, and mesenchymal tumors, and between fibrohistiocytic and leiomyocytic tumors, and therefore are of diagnostic value in histopathologic problems of the skin.

Proteoglycans detected by monoclonal antibodies in adenoid cystic carcinoma of salivary glands.

Thirty-two cases of adenoid cystic carcinoma (ACC) of the salivary glands were examined immunohistochemically by monoclonal antibodies to proteoglycans (PG) such as chondroitin 6 sulfate (C6SPG), chondrotin 4 sulfate (C4SPG), dermatan sulfate (DSPG), heparan sulfate (HSPG) and keratan sulfate (KSPG) in conjunction with specific enzymatic digestion. The cribriform structure of ACC consisted of basaloid, myoepithelium-derived, and luminal tumor cells. The myoepithelial tumor cells were positive for PG, whereas luminal tumor cells were unstained. Occasional pseudocysts also gave positive staining for PG. Tubular structures consisting of modified myoepithelial cells indicated a high intensity reaction for C6SPG and C4SPG, and a slight one for DSPG, HSPG, and KSPG. Immunodepositions in solid and cluster structures were comparatively light in terms of PG. Basement membrane in ACC stained strongly for C4SPG, slightly for C6SPG, and very slightly for DSPG, HSPG, and KSPG. In ACC, immunohistochemical staining of PG was regularly positive in myoepithelium-derived tumor cells, but was irregular in other types of tumor cells.

Pleomorphic adenoma and adenoid-cystic carcinoma of the salivary glands: comparative immunohistochemical patterns.

A series of 20 cases of pleomorphic adenoma and 19 cases of adenoid-cystic carcinoma of the salivary glands, and one case in the mammary location, were investigated regarding immunohistochemical reactivity for Tissue Polypeptide Antigen (TPA), Pre-Keratins, Vimentin, S-100 Protein, and their arrangement pattern of fibronectin. As a whole, the results support the hypothesis of morpho-structural and mainly, onto-histogenetic similarities between these tumours, but they also underline the need for great care in outlining their morpho-functional features, in relation to their different prognoses.

Malignant dermal cylindromas. Do they exist? A morphological and immunohistochemical study and review of the literature.

Malignant dermal cylindromas are very rare. We present a case of multiple cylindromas of the scalp with metastasis to a cervical lymph node. The morphology of the tumour was unusual in that it contained eccrine spiradenoma-like areas and foci of squamous differentiation with keratin formation. The immunohistochemical phenotype of the eccrine spiradenoma-like areas and the metastatic tumour was similar, but different from the areas of typical cylindroma. Although alleged "malignant" cylindromas have been reported, none have been described to have metastasized, whereas metastatic eccrine spiradenoma is well-documented. We reiterate that overlaps between dermal cylindroma and eccrine spiradenoma are more common than has been documented. In the rare event of metastases, it is the eccrine spiradenomatous component that is metastatic. We contend that there is no evidence that pure dermal cylindromas have metastasized.

Immunohistochemical identification of epithelial membrane antigen in sweat gland tumors by the use of a monoclonal antibody.

A total of 78 cases of adnexal tumors of the skin were examined with the use of monoclonal antibody against epithelial membrane antigen (EMA). The EMA reaction was confined to luminal surfaces and lateral borders of sweat glands, both eccrine and apocrine, being usually absent in ductal segments. Neoplastic lesions of all the adenomatous tumours and mixed tumours of sweat glands showed specifically positive EMA staining of luminal surfaces and lateral borders of tubular, duct-like, and cystic structures. However, solid foci of those tumours were negative for EMA. Tumours of ductal origin, e.g. syringomas and poromas, exhibited positive EMA staining in their plasma membrane, although normal intact keratinocytes did not stain for EMA. Immunohistochemical distribution of EMA in skin adnexal tumours was classified into two types: one in which the luminal surfaces and lateral outer borders were positive, similar to that of the normal secretory coil, and the other in which the plasma membrane of neoplastic cells of ductal origin was positive.

Immunolocalization of keratin proteins in sweat gland tumours by the use of monoclonal antibody.

A total of 34 cases (eccrine poroma: 2, cylindroma: 2, eccrine spiradenoma: 4, syringocystadenoma papilliferum: 1, hydroadenoma papilliferum: 1, clear cell hydroadenoma: 7, mixed tumour: 16) of sweat gland tumours of the skin were described in terms of immunohistochemical distribution of keratins using polyclonal anti-keratin antiserum (TK, detecting 41-56 KDa keratins) and monoclonal antibodies (KL1, 55-57 KDa; PKK1, 40, 45, 52.5 KDa). Keratin expression in eccrine poroma, spiradenoma and syringocystadenoma was similar to that in the ductal segment of normal sweat glands. Cylindroma showed usually slight staining for kertins. Tumour cells of hydroadenomas showed not so prominent staining for any of the keratins; however, histologically, tumour cells indicated marked variation, and the degree of keratin proteins also was different among these histological variants. Mixed tumours of the skin were strongly decorated with anti-keratin antibodies along the luminal surface cells of typical structures, while no staining occurred in outer side cells. Luminal tumour cells may be differentiated from secretory coil cells, whereas outer side cells may have a myoepithelial origin, as outer layer cells found in pleomorphic adenoma of salivary glands.

Collagenous spherulosis. A comment on its histogenesis.

Collagenous spherulosis is a benign breast lesion involving lobular acini and ductules consisting of eosinophilic spherules measuring up to 100 mu in diameter. It is a myoepithelial product. We described similar lesions in salivary gland tumors and a benign lymphoepithelial lesion of the parotid gland.

Basal membrane associated substances in human salivary glands and salivary gland tumours.

Human salivary gland tissue was analysed with respect to the distribution of basal membrane associated substances. Collagen type IV and laminin were studied on the basis of monoclonal antibodies, fibronectin was analysed with polyclonal antibodies. The structure of the basal membrane was well preserved in normal salivary gland tissue. There was a continuous staining of the basal membrane around the acini and the ducts. The labelling for these substances appeared to be associated with the myoepithelial cells. Pleomorphic adenomas exhibited a heterogeneous pattern for the basal membrane substances. Focally, there was an augmentation of collagen IV and laminin, as well as of fibronectin. This was observed in the neighbourhood of myoepithelial like cells. Other parts of pleomorphic adenomas showed an interruption of the basal membrane. Adenoid cystic carcinomas displayed a clear staining of the basal membrane associated substances in the pseudocysts. Stromal trabeculae were stained in an irregular manner. Acinic cell tumours, adenocarcinomas, mucoepidermoid tumours and squamous cell carcinomas (for comparison taken from other regions in the head and neck area) presented a clear destruction of the basal membrane as visualized by antibodies against collagen IV and laminin. The study of the basal membrane substances may be helpful for identifying special features of salivary gland tumours and for grading the amount of invasive behaviour in the malignant tumours.

Primary adenoid cystic carcinoma of the esophagus. Report of a case and its histochemical characterization.

A case of primary adenoid cystic carcinoma of the esophagus was reported. A 54-year-old male patient had a tumor in the middle third of the esophagus. Barium swallow and endoscopic examination showed a protruding tumor with a shallow ulceration in its vertex. Histological examination of the surgically removed material revealed that it was largely confined to the submucosal layer, with no metastasis to lymph nodes. The tumor cells presented three distinct patterns: globular nest with irregular cystic spaces, trabecular nest, and true tubule. Electron microscopic study revealed no microvilli or triad along cystic spaces in the globular nests. Histochemical study disclosed that cystic spaces and interstitium were rich in glycosaminoglycans. The luminal surface of the true tubules of the tumor, on the other hand, was characteristically lined by sialomucins. The trabecular type-nests differed from the globular nests in showing GSA-II reactivity and containing abundant retinol-binding protein. These results indicate that the three patterns represented different differentiations of the tumor cells. Twenty-three cases of esophageal adenoid cystic carcinoma were found in the literature from 1950 to 1983 and discussed collectively.

Immunocytochemical detection of carcinoembryonic antigen in salivary gland tumors.

The immunoperoxidase method was applied for carcinoembryonic antigen (CEA) detection in biopsy specimens of salivary gland tumors. 9 out of 10 adenoidcystic carcinomas revealed a strong and abundant reaction in tumoral glands. 10 other specimens of pleomorphic adenomas showed weak staining in the areas of epithelial proliferation. Normal glands adjacent to the tumor mass revealed a weak but constant reaction on the luminal border. As in other types of gland tumor, the quantitative estimation of CEA production by salivary gland tumors may be useful in the monitoring of recurrencies.

Immunoelectron microscope demonstration of the basement membrane components laminin and type IV collagen in the dermal cylindroma.

Specific antisera to the human laminin P1 fragment and the 7S domain of type IV collagen were used to investigate the ultrastructural location of these main basement membrane (BM) components in a dermal cylindroma. Thick frozen sections were treated by the immunoperoxidase method, postfixed, embedded in epon and then sectioned for electron microscopic examination. Laminin and type IV collagen were detected in all layers of the cylindroma BM, together with non-specifically stained, diffusely distributed fibrillar structures, possible microfibrils and anchoring fibrils or residual stromal collagen fibres. The mixed presence of the BM components indicates that these substances codistribute rather than occurring as separate layers even when forming neoplastic basement membranes.