Irving Dardick

Histogenesis of salivary gland pleomorphic adenoma (mixed tumor) with an evaluation of the role of the myoepithelial cell.

In the past a variety of cellular differentiation pathways have been proposed to account for the derivation of pleomorphic adenomas. Among these proposals, the extent of participation of the myoepithelial cell and the embryologic origin and functional role of this type of cell in this tumor have been controversial. Recent developmental, immunohistologic, and morphologic investigations suggest that the myoepithelial cell is of ectodermal origin, and a modified acinar or ductal cell possibly with a specific functional role. Autoradiographic findings indicate that reserve interaalated cells are not the principal source of new acinar or ductal cells but that both types of cells can be a self-renewing population. Thus, a neoplastically altered acinar or ductal cell, with the potential for expressing myoepithelial cell features, could be involved in the genesis of salivary gland pleomorphic adenomas. Such a hypothesis remains to be clarified. With respect to salivary gland pleomorphic adenomas, light microscopic and ultrastructural studies indicate that fully differentiated myoepithelial cells are not the principal cell found in the myxoid regions of these tumors. In these studies the major type of cell clearly has duct epithelial characteristics, frequently showing distinct evidence of squamous differentiation. In some pleomorphic adenomas, tumor cells ultrastructurally have varying degrees of epithelial characteristics with prominent intracytoplasmic microfilaments producing a myoepithelium-like appearance, whereas in other cases myxoid areas contain cells of a relatively undifferentiated type. The wide range of morphologic expression evident in pleomorphic adenoma would appear to be the result of a number of factors: the amount and distribution of mucopolysaccharide matrix formed by tumor cells in myxoid appearing areas, the expression of squamous, chondroid, or osseous metaplasia, and, ultrastructurally, the relative frequency of tumor cells with ductal, myoepithelial, myoepithelial-like, or poorly differentiated characteristics. It is proposed that this class of salivary gland tumor forms a spectrum with monomorphic adenoma and myoepithelioma at the extremes and a wide range of pleomorphic adenoma between, depending on the type and degree of gene expression that is coupled with neoplastic transformation.

Myoepithelioma: Definitions and Diagnostic Criteria

Due to their infrequency and multiplicity of histopathology, myoepitheliomas present difficulties in diagnosis and classification. Cellular varieties can be misdiagnosed as malignancies. Improvements in and clarification of diagnostic criteria are, therefore, required. A key to determining diagnostic criteria for myoepitheliomas is to study cellular morphology, cytoplasmic filament expression, and ultrastructural features of the nonluminal, i.e., neoplastic myoepithelial/basal, tumor cells of pleomorphic adenomas, and apply this information to defining myoepitheliomas. Cytologic and growth patterns of nonluminal cells in pleomorphic adenomas, including plasma-cytoid cells, are reflected in myoepitheliomas. Results also indicate that muscle-specific actin and myofilaments are expressed only in a proportion of cases, and generally in not more than 60-70% of nonluminal cells in pleomorphic adenoma; this also applies to benign and malignant myoepitheliomas. The absence of these markers does not exclude a diagnosis of myoepithelioma. Vimentin and glial acidic fibrillary protein, however, are strongly and diffusely expressed in the majority of pleomorphic adenomas and myoepitheliomas and are more reliable markers for these tumors than muscle-specific actin. Like so many other salivary gland tumors, myoepitheliomas present an equally complex histomorphology and variable expression of antigenic markers, only some of which are associated with myoepithelial and basal cells of the acini and ducts of the normal salivary gland.

Noninvasive and minimally invasive carcinoma ex mixed tumor: A clinicopathologic and ploidy study of 12 patients with major salivary tumors of low (or no?) malignant potential

We studied 12 histologically malignant salivary tumors that showed complete encapsulation or only limited microscopic invasion. Most cases were histologically characterized by atypical and mitotically active luminal cells forming dilated, angular, variably sized glands in the subcapsular region, varying proportions of nonluminal tumor cells, and a background of central fibrosed hyalinized stroma. The appearance is that of a low-grade carcinoma. Focal higher grade carcinoma was superimposed on this histologic data in three cases. Neither recurrences nor metastases were seen in 11 of 12 patients after surgical resection with a follow-up of 1.2 to 13 yrs (mean, 4.2 years). Ploidy studies were performed on the paraffin-embedded tissue in 11 cases and yielded results for 7 cases. Aneuploid cell populations were found in five tumors; two had normal diploid populations; and the ploidy results are not predictive of tumor behavior. This type of salivary gland tumor fits diagnostically within the category of noninvasive and minimally invasive carcinoma ex pleomorphic adenoma (also referred to as in situ and low-grade malignant mixed tumors), a class that requires additional awareness and precise recognition as it signifies a good prognosis after surgical resection.

Immunohistochemistry and ultrastructure of myoepithelium and modified myoepithelium of the ducts of human major salivary glands: Histogenetic implications for salivary gland tumors

The organization of salivary gland ducts, especially the presence or absence of myoepithelial cells, is central to histogenetic approaches to the classification of salivary gland tumors. Striated and excretory ducts are reported to be devoid of myoepithelial cells but do contain basal cells. To investigate the nature of such basal cells, tissue sections of normal human salivary glands were examined by means of immunohistochemical, ultrastructural, and fluorescent microscopic techniques. With the use of a mouse monoclonal anticytokeratin antibody (3 12C8-1) that, in salivary glands, is specific for myoepithelial cells, these cells associated with acini and intercalated ducts were strongly stained, as were the basal cells of striated and excretory ducts in each case. Ultrastructurally, some basal cells of both striated and excretory ducts had narrow, elongated cellular processes or the main portion of the cell containing parallel arrays of microfilaments with linear densities and micropinocytotic vesicles, whereas in other basal cells tonofilament bundles predominated. A similar range of cytoplasmic features existed in myoepithelial cells associated with acinar and intercalated duct cells. In addition, some duct basal cells have a complement of actin filaments similar to classic myoepithelium of acini and intercalated ducts. Striated and excretory ducts of human salivary glands, therefore, contain fully differentiated and modified myoepithelial cells, both of which express a specific cytokeratin polypeptide that is absent from duct luminal and acinar cells. Differentiation patterns in the intralobular and interlobular ducts suggest that these regions of salivary gland parenchyma cannot be excluded as histogenetic sites for the induction of salivary gland tumors in which neoplastic myoepithelial cells have been shown to have a major role.

Ultrastructural spectrum of hemangiopericytoma: a comparative study of fetal, adult, and neoplastic pericytes.

Since ultrastructural examination is often employed to assess controversial soft tissue tumors, it is important to be aware of the range of differentiation assumed by the tumor cells in hemangiopericytomas. For this purpose, 35 examples (10 localized to the central nervous system and 25 located peripherally) were examined ultrastructurally, and, of these, 20 cases were also studied immunohistochemically for the presence of intermediate filaments and muscle-specific actin. Based on cytologic characteristics evident by electron microscopy, tumor cell differentiation was classed as pericytic (32%), myoid (8%), nondescript (48%), fibroblastic (4%), and histiocytic (8%). Vimentin was the only intermediate filament expressed in the normal pericytes of human fetal and adult tissues and in the neoplastic pericytes of all of the hemangiopericytomas. Muscle-specific actin was present in normal pericytes, but only focally in two of the hemangiopericytomas. In various combinations basal lamina-like materials, cytoplasmic processes, cytoplasmic filaments, discrete basal lamina, and poorly formed intercellular junctions were the most frequently noted features of the tumor cells in hemangiopericytomas, whether central or peripheral, and they assist, along with the organizational relationship of tumor cells and capillaries, in distinguishing this lesion from other soft tissue sarcomas.

Salivary gland myoepithelioma variants. Histological, ultrastructural, and immunocytological features.

The histological and ultrastructural features of five major salivary gland tumours, which have little or no evidence of duct- or gland-type differentiation in routine sections, are described. Four of the cases have the tumour cells organized as narrow, anastomosing cords of cells separated by a myxoid and vascularized stroma; we have designated such lesions as reticular-type myoepitheliomas. The fifth case has a solid growth pattern and is largely composed of hyaline cells, that is, a plasmacytoid myoepithelioma. Ultrastructurally, one reticular myoepithelioma reveals myoepithelial cell differentiation with microfilament aggregates, while the other three examples are composed of modified myoepithelial cells displaying widened intercellular spaces, prominent synthesis of extracellular glycosaminoglycans, distinct basal lamina development, and obvious accumulations of cytoplasmic intermediate filaments. In electron micrographs, the modified myoepithelial cells of the plasmacytoid variant closely resemble the tumour cells in the reticular form. Three cases had expression of both glial fibrillary acid protein (GFAP) and vimentin, but only one of the myoepitheliomas contained muscle-specific actin. At least focally, each of the cases exhibited a considerable spectrum of cytokeratin filaments. Using double-labeled immunofluorescent microscopy of one reticular variant and the plasmacytoid myoepithelioma, there was individual tumour cell co-expression of GFAP and vimentin focally in the plasmacytoid myoepithelioma, but co-expression of cytokeratins 13, 16 and GFAP were not noted in either case. As expected, co-expression of high- and low-molecular weight cytokeratin filaments was wide-spread in both myoepitheliomas. Most described myoepitheliomas have a solid growth pattern and are composed of spindle and plasmacytoid cells, but based on cytological features and growth patterns in this series, it is apparent that polygonalshaped cells with novel architecture can occur in myoepitheliomas. The results also indicate the close relationship between pleomorphic adenoma and such variants of myoepithelioma.

Characterization of cytoskeletal proteins in basal cells of human parotid salivary gland ducts

From previous immunofluorescent, immunohistochemical and ultrastructural studies, myoepithelial cells have been reported to be absent from the striated and excretory ducts of human salivary gland. Yet recently, certain anti-cytokeratin monoclonal antibodies which specifically label the myoepithelium of salivary gland acini and intercalated ducts have also been found to stain basally situated cells in both striated and excretory ducts. In this study, we have used eight samples of normal human parotid gland (methacarn-fixed and frozen sections) to establish if basal cells of striated and excretory ducts have the cytoskeletal protein complement (actin and cytokeratins) of myoepithelial cells. Using a muscle-specific actin, HHF35, not only is the myoepithelium of acini and intercalated ducts stained in all cases, but stellate and spindle shaped cells are also detected all along the inter- and intralobular striated ducts in four of the eight examples. With double-labeled frozen sections and fluorescent microscopy, the actin-specific probe, phalloidin, and the myoepithelial-selective anti-cytokeratin 14 antibody, 312C8-1, confirm that the striated duct does have a population of basal cells with the cytoskeletal protein make-up of myoepithelium. The monoclonal antibody 8.12 (specific for cytokeratin 13 and 16) also stains some basal cells of striated and excretory ducts, as well as luminal cells of ducts at all levels, but does not label the myoepithelium of acini and intercalated ducts. Both the anti-cytokeratin antibodies and the actin-detecting mechanisms reveal that the basal cell population of striated and excretory ducts is more heterogeneous, and likely functionally more complex, than has been realized previously. Such findings are not in agreement with certain aspects of current theories of the histogenesis of salivary gland tumours.

Intermediate filament expression in normal parotid glands and pleomorphic adenomas.

A comparative immunohistochemical study of intermediate filament expression in normal parotid glands and pleomorphic adenomas (PA) was performed using material fixed in a modified methacarn fixative. The normal myoepithelial cells of acini stained only with monoclonal antibodies 312C8-1 (cytokeratin (CK) 14) and 4.62 (CK 19) while myoepithelial/basal cells of ducts also reacted with antibodies 8.12 (CK 13, 16), 8.60 (CK 10, 11, +1), and PKK1 (CK 7, 8, 17, 18). Normal duct luminal cells showed a different CK profile, reacting consistently with ECK, a polyclonal antibody to epidermal prekeratin (CK 3,6), and monoclonal antibodies 4.62, PKK1 and 8.60. In PA, tumour cells at the periphery of ducts, in solid areas, and at the edge of myxoid regions all had CK profiles similar to normal myoepithelial/ basal cells except that antibody 4.62 was generally negative. Vimentin and glial fibrillary acidic protein (GFAP) were uniformly negative in normal parotids but showed variable (often strong) reactivity with some cells in chondroid, myxoid and solid areas of PA. A surprising feature of most PA was the variability of CK subtype expression not only from one case to another but also within morphologically similar areas of the same specimen. These results suggest that the morphology of PA is the result of diversity of tumour cell differentiation rather than the processes implicit in a reserve cell histogenetic model.

Adenoid cystic carcinoma : Use of cell proliferation, BCL-2 expression, histologic grade, and clinical stage as predictors of clinical outcome

Although the three basic histologic growth patterns of adenoid cystic carcinomas (tubular, cribriform, and solid) provide some indication of clinical outcome, additional, perhaps superior, predictors of biologic activity are needed for patient management.

A review of the proliferative capacity of major salivary glands and the relationship to current concepts of neoplasia in salivary glands

The classification of salivary gland tumors relies heavily on histogenetic postulates. One of these, the semipluripotential reserve cell theory, suggests that certain reserve cells in specific segments of the duct system of major and minor salivary glands are critical to the development of neoplasms in these glands. However, direct evidence in support of this hypothesis is unavailable. This survey of proliferative capacity in normal salivary gland is based on a review of data in the literature, our observations of DNA synthetic and mitotic activity in developing rat and human salivary gland, and autoradiographic studies of induced cell proliferation in rat salivary gland. Autoradiography of neonatal rat salivary gland after tritiated thymidine administration, and electron microscopy of these tissues, reveals that as well as duct basal cells, luminal cells at all levels of the duct system and even acinar cells are capable of DNA synthesis and mitosis. Indeed, in such studies, more luminal than basal cells are seen in mitosis. In adult rat salivary gland induced to undergo hyperplasia, more acinar cells than intercalated duct cells are in the S phase of the cell cycle. However, cycling cells were observed even in striated ducts and, importantly, both basal and luminal cells of major interlobular excretory ducts are also labeled. Similar findings are present in fetal and adult human salivary glands. From such observations, it is evident that dividing cells are not limited to basal cells of excretory ducts and luminal cells of intercalated ducts, so that there is no support for the semipluripotential bicellular reserve cell hypothesis. However, there is considerable evidence for a multicellular theory of tumor histogenesis; that is, any of the multiplicity of cell types in normal salivary gland have the potential to give use to any of the various types of tumor occurring in this organ.