Acta histochem. (lena) 93, 402-410 (1992) Gustav Fischer Verlag Jena . Stuttgart· New York
1 2 3
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Department of Pathology, Fujian Medical College, Fuzhou, Fujian, The People's Republic of China, Department of Oral and Maxillofacial Surgery, Asahi University School of Dentistry, Hozumi, Gifu, and Department of Dermatology, Gifu University School of Medicine, Gifu, Japan
Vimentin expression in sweat gland tumours JIAN WEN HUANG I , MAYUKO KUNIKATA 2, KOUJI HASHIMURA 2, FUMINORI SAKAMOT02, KAZUMASA OGATA 2, MASAHIKO MORI2, KAZUFUMI YONEDA3, MAKOTO YANAGIHARA 3 and SHUNJI MORI3
With 5 Figures Accepted 12 April 1992
Summary Immunohistochemical localization of vimentin was studied in 93 cases of sweat gland tumours using a monoclonal anti-vimentin antibody. A strong immunoreactivity of vimentin was observed in modified myoepithelial or neoplastic myoepithelial cells of mixed tumour of the skin, syringoma, and sweat gland adenoma. Tumour cells in outer layers of tubular, ductal, and duct-like structures usually showed positive staining for vimentin, which coincided with modified myoepithelial cells. All tumour cells of clear cell hydroadenoma showed positive vimentin staining. Tumour cells of the luminal border of tubulo-ductal structures of mixed tumours were rarely immunoreactive for vimentin. Positive vimentin staining of tumour cells in the outer zone of tubulo-ductal structures in sweat gland tumours may be related to reactive proliferation of modified myoepithelial cells and simultaneous growth of luminal tumour cells.
1. Introduction Immunohistochemically detectable vimentin in mixed tumours of skin has been reported et al. 1990). Although vimentin is a marker for mesenchymal cells, recent reports have shown that vimentin is frequently co-expressed with keratin in epithelial tumour cells of salivary pleomorphic adenoma (SHINOHRARA et al. 1989; YAMADA et al. 1988), mammary tumours (GUELSTEIN et al. 1988; RAYMOND et al. 1989, DOMAGALA et al. 1990; WADA et al. 1991), kidney and renal tumours (HOLTHOFER et al. 1984; VOGEL et al. 1984), and thyroid tumours (MILTTINEN et al. 1984; UCHIDA et al. 1989). The purpose of the present study is to describe the immunohistochemical distribution of vimentin in different histologic types of sweat gland tumours using a monoclonal antibody, and to compare the results with those of salivary gland pleomorphic adenoma and mammary tumours, both of which have modified myoepithelial cells. (HASSAB-EL-NABY
2. Materials and methods 93 tumours of the sweat gland were used (Table 1). The tumour material obtained from surgery was fixed in 10% formaldehyde and embedded in paraffin. Paraffin sections were cut at 4 f-tm to detect vimentin using a monoclonal anti-vimentin antibody (mab, Dako, Denmark).
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Table I. Tumours of human sweat glands Mixed tumour of the skin Hydroadenoma Eccrine adenoma Apocrine adenoma Syringoadenoma Ecrrine spiroadenoma Hydroadenoma papilliferum Eccrine cystadenoma papilliferum Apocrine cystadenoma Ecrrine cystadenoma
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2.1. Immunohistochemical method Deparaffinized sections were first treated with methanol containing 0.3 % HzO z for 30 min, and then with normal rabbit serum (Dako, Denmark, I: 20) for 30 min. The primary antibody, mab anti-vimentin (Dako, Denmark, I :40) was applied for I h at room temperature. After washing with phosphate-buffered saline (PBS), the sections were incubated with biotinylated anti-mouse immunoglobulin (Dako, Denmark, 1: 2(0) for 30 min, followed by washing with PBS, incubation with peroxidase-conjugated streptavidin-biotin complex (ABC kit; Dako, Denmark) for 30 min, and visualization with a 0.03 % 3,3-diaminobenzidine solution containing 0.005 % HzO z. - Controls were performed by parallel incubation with PBS containing I %0 heat-inactivated bovine serum in the absence of the mab.
3. Results 3.1. Normal sweat gland No vimentin staining was present in the coiled duct, ductal segments, and myoepithelial cells of the normal eccrine and apocrine glands.
3.2. Eccrine adenoma There were great histologic variant ions such as tubular and papillary versions of tumour cell proliferation. Usually tubular structures consisted of 2 or more cell rows. Vimentin expression was restricted to the basal side of the outer cells of tubular structures, which were probably myoepithelial participant cells (Fig. 1A, B), and was confined to tumour stromal tissue (Fig. 1C, D).
3.3. Mixed tumour of the skin The tumour was composed of epithelial islets, sheets, tubules, duct-like structures and cysts, and connective tissue stroma with or without hyaline and chondroid changes. Vimentin expression was found in outer zones of basal tumour cells of tubulo-ductal epithelial structures. Outer layer cells in the ductal structures of the mixed tumour of skin showed positive vimentin staining, while luminal (inner) tumour cells were negative (Fig. 2E, F). Epithelial sheets and cords of the tumour revealed modified myoepithelial cells, which intense staining for vimentin; epithelial pearls were also stained (Fig. 3A-D). In the tumour stroma, there was strong staining of vimentin in connective tissue elements and blood vessel capillaries Z6*
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Fig. I. A: Eccrine adenoma. Tubular structures are composed of 2 cell layers, luminal epithelial cells, and basal myoepithelial cells, showing clear cytoplasm. x70. B: Ab Vimentin staining is confined to myoepithelial cells. Luminal epithelial cells are stained by a keratin antibody. x 70. C: Eccrine adenoma. Proliferation of epithelial sheets and cords into stromal connective tissue. x70. D: Ab Vimentin reaction is limited to stromal connective tissue of the tumour. x70.
(Fig. 3D). Chondroid cells markedly expressed vimentin (Fig. 3E, F). Mixed tumour of the skin were occasionally composed of modified myoepithelial cell islets and large masses containing myoepithelium-participant cells. Those modified myoepithelial cells showed eosinophilic and clear cytoplasm which were also immunoreactive for vimentin with highest to moderate degree of staining (Fig. 4A, B). Myoepithelioma-type of skin mixed tumours showed positive vimentin expression in fibrous neoplastic cells (Fig. 5 A-D).
3.4. Clear cell adenoma and hydroadenoma Clear cell or hydrolytic tumour cells were characterized by strong vimentin staining. Clear tumour cell foci were composed of cuboidal cells with strongly expressed vimentin; ductal structures lined by columnar cells also showed a positive vimentin reaction (Fig. 4C, D). Large tumour cells of clear cell hydroadenoma consisted of polygonal clear cells with round and marked basophilic nuclei in the clear cytoplasm (Fig. 4E). Vimentin staining was strongly positive in all clear tumour cells (Fig. 4F).
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Fig. 2. A: Spiroadenoma. Tubulo-duct like structure composed of luminal tumor cells showing thin epithelial cells and spindle-shaped basal cells. x 70. B: Ab Vimentin staining is restricted to basal cells, particularly abundant in basal cell cytoplasm of the basal tumour cells, and it is lacking in luminal cells. x70. C: Syringoadenoma. There are 2 cell layers in tubular structures, consisting of high columnar cells at the luminal side and small basal cells (outer side). x70. D: Ab Vimentin staining is confined to basal tumour cells, which participate in the form action of neoplastic myoepithelial cells. x 7. E: Mixed tumour of the skin; tubuloductal structures are composed of luminal cells and spindle shaped outer cells (modified myoepithelial cells) F: Outer tumour cells, spindle shaped modified myoepithelial cells of tubulo-ductal structures are stained strongly for vimentin. x70
3.5. Syringoma and syringoadenoma Typical tubular structures of the tumour were composed of 2 cell rows; columnar luminal cells and cuboidal or flattened basal cells. A marked vimentin expression was found in tumour basal cells (Fig. 2e, D).
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Fig. 3. Mixed tumour of skin, salivary gland mixed tumour type. x70. A: Irregularly-sized tubular structure consisting of proliferating modified myoepithelial cells. B: Vimentin expression in modified myoepithelial cell foci. C: Modified myoepithelial cell growth focus. D: Modified myoepithelial cells are stained strongly with anti-vimentin antibody; stromal connective tissue are also stained. E: Epithelial cords are associated with chondroidally changed foci. F: Basal parts of epithelial cord are stained for vimentin; chondroidal cells also show intense vimentin immunoreactivity.
3.6. Eccrine spiroadenoma Two types of epithelial cells were recognized, and vimentin staining was usually expressed in peripherally located tumour cells with small dark nuclei in lumina. Occasionally luminal cells stained weakly for vimentin (Fig. 2A, B).
Vimentin expression in sweat gland tumours
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Fig. 4. Myoepithelium-participant cell foci of skin mixed tumours. x 7U. A: Epithelial cell foci composed of modified myoepithelial cells. B: Vimentin staining is limited to modified myoepithelial cells. C: Large tumour mass composed of clear tumour cells corresponding to modified myoepithelial cells. D: Clear tumour cells show positive vimentin staining, and some of basally located cells are also immunoreactive. E: Clear tumour cell variation of mixed tumour of the skin. Clear tumour cells are one variation of myoepithelioma. F: The highest vimentin-staining is restricted to clear tumour cells.
Vimentin expression in sweat gland tumours was classified according to the following patterns: 1. Vimentin staining limited to stromal connective tissues. 2. Vimentin staining confined to the modified myoepithelial or neoplastic myoepitherial cells located around the tumour epithelial foci in syringoma or neoplastic myoepithelial syringoadenoma.
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Fig. 5. Myoepithelioma type of mixed skin tumours. x70. A: Solitary growth pattern of relatively fibrous type of neoplastic myoepithelial cells. B: Neoplastic myoepithelial cells react for vimentin. C: Total tumour mass is composed of modified myoepithelial cells, and tumour cells show diffuse growth. D: Tumour cells express marked vimentin staining.
3. Vimentin staining in outer tumour cells of tubulo-ductal epithelial structures, subclassified into 2 categories; firstly, vimentin in the outer side of basal tumour cells (spiroadenoma) and secondly that in tumour cells of the outer (basal) layer in syringoadenoma and mixed tumours. 4. Characteristic vimentin expression in clear tumour cells or modified myoepithelial cell foci of clear cell hydroadenomas and mixed tumours. 5. Chondroidal changed areas of mixed skin tumour characterized by strong vimentin staining.
4. Discussion Sweat glands are composed of terminal acinar secretory portions or coiled duct and ductal segments, and eccrine glands with attached hair follicles. Epithelial cells in terminal secretory portions and glandular tubules are accompanied by myoepithelial cells. Sweat gland tumours, mixed tumour, hydroadenoma, syringoma or syringoadenoma or other types of sweat gland adenomata may arise from the terminal portion of coiled duct and glandular tubes. These tumours are usually accompanied by modified myoepithelial cells or proliferating myoepithelium-participant cells. The luminal cells have been suggested to originate from terminal ductal epithelia.
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Many investigators have described the clinical behavior, histology, ultrastructure and histochemical characteristics of hydroadenomas (KEASBEY et al. 1954; O'HARA et al. 1966; WINKELMANN et al. 1967, 1968; HASHIMOTO et al. 1967). Immunohistochemically, myoepithelial cells of the sweat, salivary and mammary glands are characterized by a strongly positive immunoreactivity for actin or myosin as contractile proteins, as well as positive staining for subclasses of keratins. Myoepithelial cells in rat mammary gland have been reported as vimentin-positive (WARBURTON et al. 1989). Co-expression of vimentin and keratin has been described in developing and adult human kidney (HOLTHOFER et al. 1984), thyroid gland (MIETTINEN et al. 1984), lung alveoli, mesothelium, choroid plexus, ciliary epithelium of eye as well as in intermediate and marginal cells and perilymphatic endothelium of the inner ear of guinea-pigs (KASPER et al. 1988, 1989). Co-expression of vimentin and keratin was also found for epithelial tumour cells of salivary pleomorphic adenomas (SHINOHARA et al. 1989), adenoid cystic carcinoma (CASELITS et al. 1984; CHOMETTE et al. 1991), mammary tumours (RAYMOND et al. 1989; DOMAGALA et al. 1990; WADA et al. 1991), thyroid tumours (MIETTINEN et al. 1984; UCHIDA et al. 1989), endometrial and endocervical carcinomata (DABBSET et al. 1986), mesothelioma (LAROCCA et al. 1984), and adenocarcinomata of the lung (UPTON et al. 1986). Vimentin is a marker for mesenchymal cells and tumours of mesenchymal organ (AZUMI et al. 1987). However, recent reports have shown vimentin in epithelial tumour cells, and that vimentin does not represent a specific marker for tumours of mesenchymal origin. In the present study, vimentin-positive cells coincided in sweat gland tumours with modified myoepithelial cells or neoplastic myoepithelial cells of mixed tumours of the skin and syringoma. These immunohistochemical characteristics for vimentin expression in sweat gland tumours are similar to those of salivary pleomorphic adenomata rather than breast tumours. Vimentin staining in clear cells of hydroadenomata and clear cell variations of sweat gland adenomata with those of clear cell adenoma are comparable to those of salivary gland tumours. Recently, it has been reported that modified myoepithelial, and neoplastic myoepithelial cells express a S-100 protein in mixed tumours of skin and pleomorphic adenoma of the salivary gland (NODA et al. 1988; MORl et al. 1990). Thus myoepithelium-participant cells might show a complex pattern of multiple expression of keratin, vimentin, and S-IOO protein. The immunohistochemical analysis of the multiple expression of these markers in mixed tumour of the skin is currently being performed in our laboratory.
References AZUMI, N., and BATTIFORA, H., The distribution of vimentin and keratin in epithelial and nonepithelial neoplasmas: A comprehensive immunohistochemical study on formalin- and alcohol-fixed tumors. Am. J. Clin. Pathol. 88, 286-296 (1987). CASELlTZ, J., BECKER, J., SEIFERT, G., WEBER, K. D., and OSBORN, M., Coexpression of keratin and vimentin filaments in adenoid cystic carcinomas of salivary glands. Virchows Arch. A 403, 337-344 (1984). CHOMETTE, G., AURIOL, M., VAILLANT, J. M., KASAl, Z., OKADA, Y., and MORI, M., Heterogenesty and co-expression of intermediate filament proteins in adenoid cystic carcinoma of salivary glands. Pathol. BioI. 39, 110-116 (1991). DABBSET, D. J., GEISINGER, K. R., and NORRIS, H. T., Intermediate filaments in endometrial and endocervical carcinomas: The diagnostic utility of vimentin patterns. Amer. J. Surg. Pathol. 10, 568-576 (1986). DoMAGALA, W., LASOTA, J., DUKOWICZ, A., MARKIEWSKI, M., STRIKER, G., WEBER, K., and OSBORN, M., Vimentin expression appears to be associated with poor prognosis in node-negative ductal NOS breast carcinomas. Amer. J. Pathol. 137,1299-1304 (1990). GUELSTEIN, V. I., TCHYPYSHEVA, T. A., ERMILOVA, V. D., and LITVINVA, L. V., TROYANOVSKY, S. M., and BANNIKOV, G. A., Monoclonal antibody mapping of keratin 8 and 17 and of vimentin in normal mammary gland, benign tumours, dysplasias and breast cancer. Int. J. Cancer 42, 147-153 (1988). HASHIMOTO, K., DIBELLA, R. J., and LEVER, W. F., Clear cell hydradenoma. Histological, histochemical, and electronmicroscopic studies. Arch. Dermatol. 96, 18-38 (1967). HASSAB-EL-NABY, H. M., TAM, S., WHITE, W. L., and ACKERMAN, A. B., Mixed tumours of the skin. A histological and immunohistochemical study. Am. J. Dermatopathol. 11,413-428 (1990).
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HOLTHOFER, H., MIETTINEN, A., LEHTO, V.-P., LEHTONEN, E., and VIRTANEN, I., Expression of vimentin and cytokeratin types of intermediate fIlament proteins in developing and adult human kidneys. Lab. Invest. 50, 552-559 (1984). KASPER, M., MOLL, R., STOSIEK, P., and KARSTEN, D., Patterns of cytokeratin and vimentin expression in the human eye. Histochemistry 89,369-377 (1988). - Patterns of cytokeratinlvimentin coexpression in the guinea pig. Acta histochem. 86, 85-91 (1989). KEASBEY, L. E., and HADLEY, G. G., Clear cell hydradenoma. Report of three cases with widespread metastases. Cancer 7,934-952 (1954). LAROCCA, P.l. D., and RHEINWALD, 1. G., Coexpression of simple epithelial keratins and vimentin by human mesothelium and mesothelioma in vivo and in culture. Cancer Res. 44, 2991-2999 (1984). MIETTINEN, M., FRANSSILA, K., LEHTO, V.-P., PAASIVUO, R., and D'VIRTANEN, I., Expression of intermediate filament proteins in thyroid gland and thyroid tumors. Lab. Invest. 50, 262-270 (1984). MORl, M., YAMADA, K., TANAKA, T., and OKADA, Y., Multiple expression of keratins, vimentin, and S-Ioo protein in pleomorphic salivary adenomas. Virchows Arch. B 58, 435-444 (1990). NODA, Y., HORlKE, H., TANIMURA, T., TSUJlMURA, T., and MORI, M., Immunohistochemical localization by monoclonal antibodies of S-Ioo IX and ~ proteins in mixed tumors and adenomas of the skin. Virchows Arch. B 54,371-380 (1988). O'HARA, 1. M., BENSCH, K., IOANNIDES, G., and KLANS, S. N., Eccrine sweat gland adenoma, clear cell type. A histochemical study. Cancer 19, 1438 - 1450 (1966). RAYMOND, W. A., and LEONG, A. S., Coexpression of cytokeratin and vimentin intermediate filament proteins in benign and neoplastic breast epithelium. 1. PathoI. 157,299-306 (1989). SHINOHARA, H., YAMADA, K., TANAKA, T., MEENAGHAN, M. A., TAKAI, Y., and MORl, M., Coexpression of keratin and vimentin in salivary pleomorphic adenoma. 1. Oral PathoI. Med. 18,133-139 (1989). VOGEL, A. M., GOWN, A. M., CANGHLAN, 1., HAAS, 1. E., and BECKWlTH, 1. B., Rhabdoid tumors of the kidney contain mesenchymal specific and epithelial specific intermediate filament proteins. Lab. Invest. 50, 232-238 (1984). WADA, T., YASUTOMI, M., HASHIMURA, K., KUNlKATA, M., TANAKA, T., and MORl, M., Vimentin expression in benign and malignant lesions in human mammary gland. Anticancer Res. (in press). WARBURTON, M. 1., HUGHES, C. M., FERNS, S. A., and RUDLAND, P. S., Localization of vimentin in myoepithelial cells of the rat mammary gland. Histochem. 1. 21, 679-685 (1989). WINKELMANN, R. K., and WOLF, K., Solid cystic hydradenoma of the skin clinical and histopathologic study. Arch. DermatoI. 97,651-661 (1968). - - - Histochemistry of hydroadenoma and eccrine spiroadenoma. J. Invest. DermatoI. 49,173-180 (1967). UCHIDA, H., NAKAYAMA, I., and NOGUCHI, S., An immunohistochemical study of cytokeratin and vimentin in benign and malignant thyroid lesions. Acta PathoI. lap. 39,169-175 (1987). UPTON, M. P., HIROHASHI, S., TOME, Y., MIYAZAWA, N., SUEMASU, K., and SHIMOSATO, Y., Expression of vimentin in surgically resected adenocarcinomas and large cell 'carcinomas of lung. Am. 1. Surg. PathoI. 10, 560-567 (1986). YAMADA, K., SHINOHARA, H., TAKAI, Y., and MaRl, M., Monoclonal antibody-detected vimentin distribution in pleomorphic adenoma of salivary glands. J. Oral PathoI. 17,348-353 (1988). Author's address: Dr. 1. W. HUANG, Department of Pathology, Fujian Medical College, Fuzhou, Fujian, The People's Republic of China.
1 2 3
"lda
hiSltthe.ia
Department of Pathology, Fujian Medical College, Fuzhou, Fujian, The People's Republic of China, Department of Oral and Maxillofacial Surgery, Asahi University School of Dentistry, Hozumi, Gifu, and Department of Dermatology, Gifu University School of Medicine, Gifu, Japan
Vimentin expression in sweat gland tumours JIAN WEN HUANG I , MAYUKO KUNIKATA 2, KOUJI HASHIMURA 2, FUMINORI SAKAMOT02, KAZUMASA OGATA 2, MASAHIKO MORI2, KAZUFUMI YONEDA3, MAKOTO YANAGIHARA 3 and SHUNJI MORI3
With 5 Figures Accepted 12 April 1992
Summary Immunohistochemical localization of vimentin was studied in 93 cases of sweat gland tumours using a monoclonal anti-vimentin antibody. A strong immunoreactivity of vimentin was observed in modified myoepithelial or neoplastic myoepithelial cells of mixed tumour of the skin, syringoma, and sweat gland adenoma. Tumour cells in outer layers of tubular, ductal, and duct-like structures usually showed positive staining for vimentin, which coincided with modified myoepithelial cells. All tumour cells of clear cell hydroadenoma showed positive vimentin staining. Tumour cells of the luminal border of tubulo-ductal structures of mixed tumours were rarely immunoreactive for vimentin. Positive vimentin staining of tumour cells in the outer zone of tubulo-ductal structures in sweat gland tumours may be related to reactive proliferation of modified myoepithelial cells and simultaneous growth of luminal tumour cells.
1. Introduction Immunohistochemically detectable vimentin in mixed tumours of skin has been reported et al. 1990). Although vimentin is a marker for mesenchymal cells, recent reports have shown that vimentin is frequently co-expressed with keratin in epithelial tumour cells of salivary pleomorphic adenoma (SHINOHRARA et al. 1989; YAMADA et al. 1988), mammary tumours (GUELSTEIN et al. 1988; RAYMOND et al. 1989, DOMAGALA et al. 1990; WADA et al. 1991), kidney and renal tumours (HOLTHOFER et al. 1984; VOGEL et al. 1984), and thyroid tumours (MILTTINEN et al. 1984; UCHIDA et al. 1989). The purpose of the present study is to describe the immunohistochemical distribution of vimentin in different histologic types of sweat gland tumours using a monoclonal antibody, and to compare the results with those of salivary gland pleomorphic adenoma and mammary tumours, both of which have modified myoepithelial cells. (HASSAB-EL-NABY
2. Materials and methods 93 tumours of the sweat gland were used (Table 1). The tumour material obtained from surgery was fixed in 10% formaldehyde and embedded in paraffin. Paraffin sections were cut at 4 f-tm to detect vimentin using a monoclonal anti-vimentin antibody (mab, Dako, Denmark).
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Table I. Tumours of human sweat glands Mixed tumour of the skin Hydroadenoma Eccrine adenoma Apocrine adenoma Syringoadenoma Ecrrine spiroadenoma Hydroadenoma papilliferum Eccrine cystadenoma papilliferum Apocrine cystadenoma Ecrrine cystadenoma
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2.1. Immunohistochemical method Deparaffinized sections were first treated with methanol containing 0.3 % HzO z for 30 min, and then with normal rabbit serum (Dako, Denmark, I: 20) for 30 min. The primary antibody, mab anti-vimentin (Dako, Denmark, I :40) was applied for I h at room temperature. After washing with phosphate-buffered saline (PBS), the sections were incubated with biotinylated anti-mouse immunoglobulin (Dako, Denmark, 1: 2(0) for 30 min, followed by washing with PBS, incubation with peroxidase-conjugated streptavidin-biotin complex (ABC kit; Dako, Denmark) for 30 min, and visualization with a 0.03 % 3,3-diaminobenzidine solution containing 0.005 % HzO z. - Controls were performed by parallel incubation with PBS containing I %0 heat-inactivated bovine serum in the absence of the mab.
3. Results 3.1. Normal sweat gland No vimentin staining was present in the coiled duct, ductal segments, and myoepithelial cells of the normal eccrine and apocrine glands.
3.2. Eccrine adenoma There were great histologic variant ions such as tubular and papillary versions of tumour cell proliferation. Usually tubular structures consisted of 2 or more cell rows. Vimentin expression was restricted to the basal side of the outer cells of tubular structures, which were probably myoepithelial participant cells (Fig. 1A, B), and was confined to tumour stromal tissue (Fig. 1C, D).
3.3. Mixed tumour of the skin The tumour was composed of epithelial islets, sheets, tubules, duct-like structures and cysts, and connective tissue stroma with or without hyaline and chondroid changes. Vimentin expression was found in outer zones of basal tumour cells of tubulo-ductal epithelial structures. Outer layer cells in the ductal structures of the mixed tumour of skin showed positive vimentin staining, while luminal (inner) tumour cells were negative (Fig. 2E, F). Epithelial sheets and cords of the tumour revealed modified myoepithelial cells, which intense staining for vimentin; epithelial pearls were also stained (Fig. 3A-D). In the tumour stroma, there was strong staining of vimentin in connective tissue elements and blood vessel capillaries Z6*
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Fig. I. A: Eccrine adenoma. Tubular structures are composed of 2 cell layers, luminal epithelial cells, and basal myoepithelial cells, showing clear cytoplasm. x70. B: Ab Vimentin staining is confined to myoepithelial cells. Luminal epithelial cells are stained by a keratin antibody. x 70. C: Eccrine adenoma. Proliferation of epithelial sheets and cords into stromal connective tissue. x70. D: Ab Vimentin reaction is limited to stromal connective tissue of the tumour. x70.
(Fig. 3D). Chondroid cells markedly expressed vimentin (Fig. 3E, F). Mixed tumour of the skin were occasionally composed of modified myoepithelial cell islets and large masses containing myoepithelium-participant cells. Those modified myoepithelial cells showed eosinophilic and clear cytoplasm which were also immunoreactive for vimentin with highest to moderate degree of staining (Fig. 4A, B). Myoepithelioma-type of skin mixed tumours showed positive vimentin expression in fibrous neoplastic cells (Fig. 5 A-D).
3.4. Clear cell adenoma and hydroadenoma Clear cell or hydrolytic tumour cells were characterized by strong vimentin staining. Clear tumour cell foci were composed of cuboidal cells with strongly expressed vimentin; ductal structures lined by columnar cells also showed a positive vimentin reaction (Fig. 4C, D). Large tumour cells of clear cell hydroadenoma consisted of polygonal clear cells with round and marked basophilic nuclei in the clear cytoplasm (Fig. 4E). Vimentin staining was strongly positive in all clear tumour cells (Fig. 4F).
Vimentin expression in sweat gland tumours
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Fig. 2. A: Spiroadenoma. Tubulo-duct like structure composed of luminal tumor cells showing thin epithelial cells and spindle-shaped basal cells. x 70. B: Ab Vimentin staining is restricted to basal cells, particularly abundant in basal cell cytoplasm of the basal tumour cells, and it is lacking in luminal cells. x70. C: Syringoadenoma. There are 2 cell layers in tubular structures, consisting of high columnar cells at the luminal side and small basal cells (outer side). x70. D: Ab Vimentin staining is confined to basal tumour cells, which participate in the form action of neoplastic myoepithelial cells. x 7. E: Mixed tumour of the skin; tubuloductal structures are composed of luminal cells and spindle shaped outer cells (modified myoepithelial cells) F: Outer tumour cells, spindle shaped modified myoepithelial cells of tubulo-ductal structures are stained strongly for vimentin. x70
3.5. Syringoma and syringoadenoma Typical tubular structures of the tumour were composed of 2 cell rows; columnar luminal cells and cuboidal or flattened basal cells. A marked vimentin expression was found in tumour basal cells (Fig. 2e, D).
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Fig. 3. Mixed tumour of skin, salivary gland mixed tumour type. x70. A: Irregularly-sized tubular structure consisting of proliferating modified myoepithelial cells. B: Vimentin expression in modified myoepithelial cell foci. C: Modified myoepithelial cell growth focus. D: Modified myoepithelial cells are stained strongly with anti-vimentin antibody; stromal connective tissue are also stained. E: Epithelial cords are associated with chondroidally changed foci. F: Basal parts of epithelial cord are stained for vimentin; chondroidal cells also show intense vimentin immunoreactivity.
3.6. Eccrine spiroadenoma Two types of epithelial cells were recognized, and vimentin staining was usually expressed in peripherally located tumour cells with small dark nuclei in lumina. Occasionally luminal cells stained weakly for vimentin (Fig. 2A, B).
Vimentin expression in sweat gland tumours
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Fig. 4. Myoepithelium-participant cell foci of skin mixed tumours. x 7U. A: Epithelial cell foci composed of modified myoepithelial cells. B: Vimentin staining is limited to modified myoepithelial cells. C: Large tumour mass composed of clear tumour cells corresponding to modified myoepithelial cells. D: Clear tumour cells show positive vimentin staining, and some of basally located cells are also immunoreactive. E: Clear tumour cell variation of mixed tumour of the skin. Clear tumour cells are one variation of myoepithelioma. F: The highest vimentin-staining is restricted to clear tumour cells.
Vimentin expression in sweat gland tumours was classified according to the following patterns: 1. Vimentin staining limited to stromal connective tissues. 2. Vimentin staining confined to the modified myoepithelial or neoplastic myoepitherial cells located around the tumour epithelial foci in syringoma or neoplastic myoepithelial syringoadenoma.
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Fig. 5. Myoepithelioma type of mixed skin tumours. x70. A: Solitary growth pattern of relatively fibrous type of neoplastic myoepithelial cells. B: Neoplastic myoepithelial cells react for vimentin. C: Total tumour mass is composed of modified myoepithelial cells, and tumour cells show diffuse growth. D: Tumour cells express marked vimentin staining.
3. Vimentin staining in outer tumour cells of tubulo-ductal epithelial structures, subclassified into 2 categories; firstly, vimentin in the outer side of basal tumour cells (spiroadenoma) and secondly that in tumour cells of the outer (basal) layer in syringoadenoma and mixed tumours. 4. Characteristic vimentin expression in clear tumour cells or modified myoepithelial cell foci of clear cell hydroadenomas and mixed tumours. 5. Chondroidal changed areas of mixed skin tumour characterized by strong vimentin staining.
4. Discussion Sweat glands are composed of terminal acinar secretory portions or coiled duct and ductal segments, and eccrine glands with attached hair follicles. Epithelial cells in terminal secretory portions and glandular tubules are accompanied by myoepithelial cells. Sweat gland tumours, mixed tumour, hydroadenoma, syringoma or syringoadenoma or other types of sweat gland adenomata may arise from the terminal portion of coiled duct and glandular tubes. These tumours are usually accompanied by modified myoepithelial cells or proliferating myoepithelium-participant cells. The luminal cells have been suggested to originate from terminal ductal epithelia.
Vimentin expression in sweat gland tumours
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Many investigators have described the clinical behavior, histology, ultrastructure and histochemical characteristics of hydroadenomas (KEASBEY et al. 1954; O'HARA et al. 1966; WINKELMANN et al. 1967, 1968; HASHIMOTO et al. 1967). Immunohistochemically, myoepithelial cells of the sweat, salivary and mammary glands are characterized by a strongly positive immunoreactivity for actin or myosin as contractile proteins, as well as positive staining for subclasses of keratins. Myoepithelial cells in rat mammary gland have been reported as vimentin-positive (WARBURTON et al. 1989). Co-expression of vimentin and keratin has been described in developing and adult human kidney (HOLTHOFER et al. 1984), thyroid gland (MIETTINEN et al. 1984), lung alveoli, mesothelium, choroid plexus, ciliary epithelium of eye as well as in intermediate and marginal cells and perilymphatic endothelium of the inner ear of guinea-pigs (KASPER et al. 1988, 1989). Co-expression of vimentin and keratin was also found for epithelial tumour cells of salivary pleomorphic adenomas (SHINOHARA et al. 1989), adenoid cystic carcinoma (CASELITS et al. 1984; CHOMETTE et al. 1991), mammary tumours (RAYMOND et al. 1989; DOMAGALA et al. 1990; WADA et al. 1991), thyroid tumours (MIETTINEN et al. 1984; UCHIDA et al. 1989), endometrial and endocervical carcinomata (DABBSET et al. 1986), mesothelioma (LAROCCA et al. 1984), and adenocarcinomata of the lung (UPTON et al. 1986). Vimentin is a marker for mesenchymal cells and tumours of mesenchymal organ (AZUMI et al. 1987). However, recent reports have shown vimentin in epithelial tumour cells, and that vimentin does not represent a specific marker for tumours of mesenchymal origin. In the present study, vimentin-positive cells coincided in sweat gland tumours with modified myoepithelial cells or neoplastic myoepithelial cells of mixed tumours of the skin and syringoma. These immunohistochemical characteristics for vimentin expression in sweat gland tumours are similar to those of salivary pleomorphic adenomata rather than breast tumours. Vimentin staining in clear cells of hydroadenomata and clear cell variations of sweat gland adenomata with those of clear cell adenoma are comparable to those of salivary gland tumours. Recently, it has been reported that modified myoepithelial, and neoplastic myoepithelial cells express a S-100 protein in mixed tumours of skin and pleomorphic adenoma of the salivary gland (NODA et al. 1988; MORl et al. 1990). Thus myoepithelium-participant cells might show a complex pattern of multiple expression of keratin, vimentin, and S-IOO protein. The immunohistochemical analysis of the multiple expression of these markers in mixed tumour of the skin is currently being performed in our laboratory.
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