Immunohistochemical localization of members of the transforming growth factor (TGF)-β superfamily in normal human salivary glands and pleomorphic adenomas

Although pleomorphic adenoma is the most common type of salivary gland epithelial tumor, it frequently contains "mesenchymal"-like components, including myxoid or chondroid tissues. We reported previously that chondroid tissue formation in pleomorphic adenoma was associated with overexpression of bone morphogenetic proteins (BMPs) by neoplastic myoepithelial cells. BMPs belong to the transforming growth factor (TGF)-beta superfamily, so we hypothesized that pleomorphic adenoma may express TGF-betas and that these molecules may regulate mesenchymal-like tissue formation. To evaluate this hypothesis, we immunohistochemically examined TGF-beta1, -beta2 and -beta3 expression and localization in normal salivary glands and in 43 cases of pleomorphic adenomas. There was no evidence of TGF-beta1 expression in normal salivary glands or pleomorphic adenomas. Signals for TGF-beta2 in the normal salivary glands were observed in the intercalated ducts, while in pleomorphic adenomas they were observed in the inner ductal cells of the tubulo-glandular structures. Signals for TGF-beta3 in the normal salivary glands were observed in mucous cells, whereas in pleomorphic adenomas they were observed in the solid nests of neoplastic myoepithelial cells, in the portion showing squamous metaplasia, and in the inner ductal cells of tubulo-glandular structures. TGF-betas induce ectopic cartilage formation in vivo, but chondroid tissues in pleomorphic adenomas showed only weak TGF-beta3 expression. TGF-beta may be related to differentiation of the inner ductal cells and the neoplastic myoepithelial cells. In conclusion, pleomorphic adenomas expressed TGF-beta2 and -beta3, which may be associated with differentiation of the inner ductal cells and neoplastic myoepithelial cells.     

Immunocytochemical localization of bone morphogenetic proteins (BMPs) in salivary gland pleomorphic adenoma

The localization of bone morphogenetic protein (BMP)-1, -2, -3 and transforming growth factor (TGF)-β in normal salivary gland and pleomorphic adenoma of the salivary gland has been examined immunocytochemically. Tumor cells with BMP immunostaining in pleomorphic adenoma were associated with some solid cellular and tubuloglandular patterns, and with stellate cells in the myxoid area. In addition, in the chondroid area of three pleomorphic adenomas, chondrocyte-like cells were positive for BMPs. It is speculated that BMPs secreted by the tumor cells play a role in the formation of the chondroid component in pleomorphic adenoma by inducing some tumor cells, probably neoplastic myoepithelial cells, to differentiate to chondrocytes by metaplastic change. No tumor cells specifically immunostained with TGF-β were found. TGF-β was positive in fibrous and hyalinized stroma. In the submandibular gland, only anti-BMP-1 antibody specifically reacted to apical portions of degenerated serous acinar cells.     

Assessment of p63 expression in the salivary gland neoplasms adenoid cystic carcinoma, polymorphous low-grade adenocarcinoma, and basal cell and canalicular adenomas

PURPOSE: The purpose of this study was to determine the extent of p63 immunoreactivity in the malignant salivary gland neoplasms adenoid cystic carcinoma (ACC) and polymorphous low-grade adenocarcinoma (PLGA) and to compare this to the expression of this marker in the benign salivary gland tumors canalicular adenoma and basal cell adenoma. Few studies on the expression of p63 in head and neck salivary gland tumors have been published to date. P63, a selective immunohistochemical marker of basal/stem cells of stratified epithelium and of myoepithelial cells, is a p53 homologue that plays an essential role in both morphogenesis of epidermis and limb development. P63 immunoreactivity has been demonstrated in squamous cell and urothelial carcinomas. It is generally absent in most nonsquamous cell carcinomas.Study design Formalin-fixed paraffin-embedded sections from 49 salivary gland neoplasms, representing 6 canalicular adenomas, 11 basal cell adenomas, 17 PLGA and 15 ACC accessioned from 1989 to 2002 by the Department of Pathology, Long Island Jewish Medical Center, New Hyde Park, NY, were stained with an anti-p63 monoclonal antibody. RESULTS: Nuclear p63 reactivity was uniformly positive in PLGA (17/17, 100%). Positive reactivity was also identified in the majority of cases of ACC (13/15, 87%), primarily in the nonluminal myoepithelial-like cells surrounding luminal cells. Canalicular adenoma did not exhibit any p63 immunoreactivity. All basal cell adenomas of parotid origin stained strongly for p63, with staining localized to the peripheral tumor cells situated adjacent to the connective tissue stroma. None of the basal cell adenomas originating in the upper lip stained with p63. In native adjacent salivary gland tissue, p63 reactivity was identified focally in the nuclei of myoepithelial and basal duct cells. CONCLUSIONS: P63 is strongly expressed in basal cell adenoma of parotid origin, and in ACC and PLGA. Canalicular adenoma did not demonstrate p63 staining, consistent with this tumor's putative luminal ductal cell differentiation. Our results suggest that the neoplastic cells in PLGA may represent either a population of p63-positive epithelial stem/reserve cells similar to the basal cells of stratified epithelium, or modified myoepithelial cells. Given the staining pattern of the tumors examined, p63 does not appear to be an ideal marker for distinguishing between ACC, PLGA, and basal cell adenoma.     

涎腺多形性腺瘤的纤维粘连素和层粘蛋白的表达和分布

纤维粘连素（ＦｉｂｒｏｎｅｃｔｉｎＦＮ）是细胞外基质蛋白；层粘蛋白（ＬａｍｉｎｉｎＬＭ）是基底膜成分，两者均对细胞的生长、分化、极向化和移动等行为起重要介导作用，本文采用免疫组织化学方法观察ＦＮ和ＬＭ在正常涎腺及涎腺多形性腺瘤的分布和表达，正常腺体中ＦＮ的表达定位于闰管细胞胞浆；ＬＭ定位于各级导管细胞胞浆，导管及腺泡的基底膜，多形性腺瘤中ＦＮ阳性细胞分散在肿瘤上皮结构区，在玻璃样、粘液样和软骨样间质中的变异肌上皮细胞ＦＮ呈强阳性，ＬＭ阳性见于肿瘤上皮细胞的邻接面和玻璃样间质中的变异肌上皮细胞，软骨样间质ＬＭ阴性     

Are myoepithelial cells responsible for the widespread expression of inducible nitric oxide synthase in pleomorphic adenoma? An immunohistochemical study

Many of the actions of nitric oxide (NO) are still poorly understood. Recently, it has been shown that the inducible isoform of the enzyme nitric oxide synthase, iNOS, is expressed in both salivary ducts and pleomorphic adenoma. The current immunohistochemistry study determined whether or not this distribution correlated with smooth muscle actin (SMA) expression, thereby suggesting the expression by myoepithelial cells in both sites. Twenty cases of histologically confirmed pleomorphic adenoma, the sections of which contained adjacent normal salivary gland tissue, were stained for iNOS and smooth muscle actin (clone 1A4). The salivary ducts of all cases were stained intensely by both antibodies, with smooth muscle actin staining also being noted around acini in the normal gland parenchyma. Moderate or heavy staining for iNOS was found in all specimens of pleomorphic adenoma, with smooth muscle actin being distributed in a similar manner in 19 cases. Smooth muscle actin, but not iNOS, was also noted in blood vessels of both normal glands and tumours. The correlation between iNOS and SMA in pleomorphic adenoma was significant (P<0.001). The presence of iNOS in normal salivary ducts and pleomorphic adenoma is most likely due to expression by myoepithelial cells.     

谷胱甘肽过氧化物酶-3在腮腺多形性腺瘤中的表达及意义

目的 :研究谷胱甘肽过氧化物酶-3(glutathione peroxidase-3,GPX-3)基因及蛋白在腮腺良、恶性多形性腺瘤中的表达,以明确GPX-3与腮腺多形性腺瘤发生、发展的相关性,为临床预测腮腺多形性腺瘤的发生及恶变提供理论依据。方法:采用荧光定量PCR和蛋白免疫印迹法检测30例腮腺多形性腺瘤,30例腮腺多形性腺瘤的瘤旁2 cm腺体组织,以及10例恶性腮腺多形性腺瘤中的GPX-3 mRNA及蛋白的表达,对其相对表达量进行统计学分析。结果:GPX-3 mRNA和蛋白在腮腺良、恶性多形性腺瘤中的表达明显低于瘤旁腺体组织,且在腮腺恶性多形性腺瘤中的表达最低,差异有统计学意义(P<0.01)。GPX-3在腮腺多形性腺瘤中的表达与患者的年龄、性别及肿瘤大小无关,差异无统计学意义(P>0.05),且GPX-3在腮腺多形性腺瘤中的表达与肿瘤TNM分期及恶性成分比例有关,其差异有统计学意义(P<0.05)。结论:GPX-3在腮腺良性多形性腺瘤中低表达,且在腮腺恶性多形性腺瘤中的表达最低,提示GPX-3的低表达与腮腺多形性腺瘤的发生及恶变密切相关。     

Expression of CD44 standard and variant isoforms in parotid gland and parotid gland tumours

In this study, we investigated the distribution of the standard form of the CD44 (CD44s) cell adhesion molecule and of its v3 and v6 isoforms in samples of foetal and adult parotid gland tissue, in comparison with samples of parotid gland adenomas and carcinoma ex pleomorphic adenoma. Foetal parotid gland showed CD44s and CD44v3 expression in the peripheral small primordial ducts and acini, while CD44v6 was only focally expressed. Adult parotid gland tissue showed a similar distribution of CD44s and variants, with a predominant expression in acinar structures and a weaker expression at duct level. In parotid gland adenomas, a diffuse and intense expression of CD44s and variants 3 and 6 was observed only in pleomorphic adenomas, while expression of CD44s was prevalent in Warthin's tumour, myoepithelioma and oncocytoma. The malignant areas of carcinoma ex pleomorphic adenoma showed a markedly decreased expression of CD44v3 and CD44v6 in comparison with the adjacent pleomorphic adenoma component. In conclusion, the prevalent expression of CD44s and variants in pleomorphic adenoma in comparison with other adenomas may be related to the abundant extracellular matrix production present in these tumours, while loss of CD44v3 and CD44v6 associated with the onset of carcinoma ex pleomorphic adenoma could promote stromal invasion, eventually contributing to the development of distant metastases.     

Immunohistochemical localization of fibroblast growth factors (FGFs) and FGF receptor-1 in human normal salivary glands and pleomorphic adenomas

Basic and acidic fibroblast growth factors (bFGF and aFGF) are heparin-binding growth factors, and promote fibrogenesis and angiogenesis. We investigated the immunohistochemical localization of bFGF, aFGF, and FGF receptor-1 in pleomorphic adenomas. In the normal salivary glands, bFGF was localized in the basement membranes of intercalated ducts, acini and basal cells of the excretory ducts, while aFGF was localized focally in the intercalated ductal cells and basal cells of the excretory ducts. In pleomorphic adenomas, bFGF was immunolocalized in the basement membranes around the solid nests of myoepithelial cells, around the neoplastic myoepithelial cells in the myxoid areas, and in the lacuna cells in the chondroid areas. In contrast, chondroid areas exhibited no immunoreactivity with aFGF. Positive signals for aFGF were localized in luminal cells of the tubuloglandular structures in pleomorphic adenomas. FGF receptor-1 immunolocalized in the lacuna cells and myoepithelial cells in the solid and myxoid areas. These observations suggest that bFGF and FGF receptor-1 produced by myoepithelial cells inhibited terminal differentiation and enchondral ossification in pleomorphic adenomas. These results also suggest important roles for FGFs in the formation of various structures with mesenchymal-like histology.     

Gli2基因在涎腺多形性腺瘤中的表达及其意义

目的:探讨Gli2基因在涎腺多形性腺瘤和正常涎腺组织中的表达水平及其在多形性腺瘤发生发展中的作用。方法:收集20例腮腺多形性腺瘤组织和20例相对应的瘤旁正常腮腺组织,应用逆转录聚合酶链反应(RT-PCR)和实时荧光定量PCR(Real-time PCR)方法检测Gli2基因mRNA的表达水平,分析其表达变化,并探讨Gli2基因在涎腺多形性腺瘤发生发展中的作用及其临床应用价值。结果:与正常腮腺组织相比,多形性腺瘤组织中Gli2基因mRNA的表达水平升高。结论:多形性腺瘤中Gli2 mRNA的高表达可诱导肿瘤细胞持续增殖,提示Gli2基因可能在涎腺多形性腺瘤的发生发展中具有重要作用。     

S-100蛋白和中间丝在涎腺多形性腺瘤中的表达

目的 观察Ｓ－１００Ａ１、Ｓ－１００Ａ４、Ｓ－１００Ａ６、Ｓ－１００Ｂ、Ｋ８．１２、ＫＬ１、波形蛋白（Ｖｉｍｅｎｔｉｎ）、胶质纤维酸性蛋白（ＧＦＡＰ）和神经元特异性烯醇化酶（ＮＳＥ）在涎腺多形性腺瘤中的表达，探讨Ｓ－１００蛋白新亚型的分布规律和作用机制以及肿瘤性肌上皮细胞的生物学特性。方法 将２３例正常涎腺和６０例涎腺多形性腺瘤常规石蜡包埋，制成４μｍ厚的连续切片，行ＨＥ和免疫组化染色。结果 在涎     

涎腺多形性腺瘤中赖氨酰氧化酶表达的研究

目的 检测涎腺多形性腺瘤(salivary pleomorphic adenoma,SPA)组织中赖氨酰氧化酶(lysyl oxidase,LOX)的表达,探讨LOX与SPA发生发展的关系.方法 取20例SPA组织标本,并配对20例SPA瘤旁涎腺组织,采用反转录聚合酶链式反应(RT-PCR)和实时荧光定量PCR方法检测LOX mRNA在组织中的的表达水平,并分析其表达变化,用蛋白质印迹法检测LOX的蛋白表达水平.结果 SPA组织中LOX mRNA表达水平明显高于瘤旁涎腺组织(12.81 ±0.92),SPA组织中LOX蛋白表达水平明显高于瘤旁涎腺组织.结论 LOX mRNA和蛋白在SPA瘤组织中均呈高表达,提示LOX的异常表达可能参与了SPA的发生、发展.     

Immunohistochemical demonstration of bone morphogenetic protein-2 and type II collagen in pleomorphic adenoma of salivary glands

Immunohistochemical investigation of bone morphogenetic protein-2 (BMP-2) and type II collagen, two cartilage-associated proteins, was undertaken using monoclonal antibodies in 20 cases of salivary pleomorphic adenoma (PA) in order to explore their possible roles in chondroid differentiation of this tumor. Other salivary gland tumors, including adenoid cystic carcinoma (17 cases), polymorphous low-grade adenocarcinoma (10 cases), basal cell adenoma (3 cases), basal cell adenocarcinoma (1 case), and epithelial-myoepithelial carcinoma (2 cases), were also examined for comparison. In PA, BMP-2 immunoreactivity was detected in the luminal and non-luminal cells of the tubulo-ductal structures, plasmacytoid cells, and other scattered tumor cells in solid areas. In addition, tumor cells in chondroid areas in most cases (14/15), and stellate cells in myxoid areas in many cases (7/19), were also intensely labeled for BMP-2. Furthermore, BMP-2 was also detected in the non-neoplastic ductal cells in salivary glands, whereas no other salivary gland tumors were positively stained for this protein. Type II collagen was localized in the intercellular matrix of chondroid areas and in a few chondroid differentiating cells in myxoid areas, confirming its cartilage-specificity. A proportional relationship was observed between BMP-2 expression and chondroid formation, although BMP-2 was also stained in occasional PAs without chondroid formation. It is speculated that BMP-2 might be secreted by tumor cells and play a role in chondroid formation in PA by inducing some tumor cells to produce type II collagen and other chondroid matrical substances, like glycosaminoglycans. The expression of BMP-2 is specific to PA and may possibly be used as a useful marker in differentiating PA from other salivary gland tumors.     

Simultaneous palatal pleomorphic adenoma and mandibular keratocystic odontogenic tumor in a pediatric patient

BACKGROUND: Neoplasms of the salivary gland account for 3% of all head and neck tumors; most of these occur in the parotid gland and are benign; 80% of them are pleomorphic adenomas. Benign neoplasms of the salivary gland are rare in children compared with adults. Keratocystic odontogenic tumor (KCOT) is considered the second most frequent neoplasm of all odontogenic tumors after the ameloblastoma. In the present report, we introduce the unusual presentation of a child with a simultaneous pleomorphic adenoma of the right hard palate and a KCOT of the left mandibular angle. DISCUSSION: The case is significant for 2 reasons; according to the literature, there are few reports of pleomorphic adenoma arising from a minor salivary gland of the palate in a patient younger than 18 years. This is to our knowledge the first recorded instance of simultaneous palatal pleomorphic adenoma and mandibular KCOT occurring in a child.     

人涎腺多形性腺瘤细胞体外培养及其生物学特性

目的 建立人涎腺多形性腺瘤细胞系。方法 取自腮腺多形性腺瘤患者的原发肿瘤进行体外培养。对培养的细胞进行活细胞观察和组织学染色。观察细胞生长状况，绘制生长曲线。采用免疫细胞化学检测细胞内特异性抗原标记物对细胞进行鏊定，并对肿瘤的致瘤性进行裸鼠异种移植实验。结果 多形性腺瘤体外培养细胞呈多边形或梭形，有的细胞围成腺腔结构，胞浆内富含粘液，形成空泡。有的成片排列，细胞外有粘液样物质。细胞的生长曲线较平缓，无明显快速生长期。肿瘤性腺上皮细胞角蛋白阳性，肿瘤性肌上皮细胞肌动蛋白阳性。结论 建立的涎腺多形性腺瘤有限细胞系(SPA-02)，保留了原代细胞的特点，并与原发肿瘤的组织学特征相似。     

PCNA, Ki-67 and p53 expressions in submandibular salivary gland tumours

Salivary gland tumours are uncommon with a broad heterogeneity. The most common benign tumour is the pleomorphic adenoma, whereas mucoepidermoid carcinoma and adenoid cystic carcinoma predominate among the malignancies. Most salivary gland tumours occur in the parotid, and consequently clinical and biological data are normally derived from this site. This work describes the expressions of PCNA, Ki-67 and p53 in 15 pleomorphic adenomas, 15 mucoepidermoid carcinomas and 15 adenoid cystic carcinomas of the submandibular gland. Our results showed that all pleomorphic adenomas were negative for p53 and Ki-67 with 66.6% being positive for PCNA. Conversely, p53 was positive in 53% of the mucoepidermoid carcinomas and in 20% of the adenoid cystic carcinomas. Ki-67 was expressed in 47.7% of the mucoepidermoid carcinomas and 40% of the adenoid cystic carcinomas. All malignant tumours were positive for PCNA. These results indicate that the proliferative rate analysed with PCNA and Ki-67 and the expression of p53 in pleomorphic adenoma and adenoid cystic carcinoma of the submandibular gland were similar to those described in the parotid and minor salivary glands. However, mucoepidermoid carcinomas showed higher expression of these markers than those of other salivary glands. This work is the first describing the expression of these immunohistochemical markers exclusively in submandibular salivary gland tumours.     

Pleomorphic Adenoma (Benign Mixed Tumour) of the Minor Salivary Glands of the Upper Lip

Pleomorphic adenoma is the benign tumor of salivary glands, which originates from the myoepithelial cells and intercalated duct cells. This tumor is more common in major salivary glands. This case report describes a rare and unusual lesion in a 55-year-old female, which was diagnosed as pleomorphic adenoma of the minor salivary glands in the upper lip. The tumor was a circumscribed, submucosal nodule, about 2.0 cm in diameter and was characterized by slow growth and rubbery consistency. Complete excision was performed and the histopathological analysis showed an epithelial salivary gland tumor with islands of plasmacytoid cells, duct like structures, in a variable stroma with chondroid, fibrous and myxoid appearance. No recurrence was observed 1 year after the surgery.     

Expression of podoplanin in the mouse salivary glands

OBJECTIVE: Podoplanin is one of the most highly expressed lymphatic-specific genes. Here, we report the distribution of cells expressing podoplanin in mouse salivary glands. DESIGN: We immunohistochemically investigated the distribution of cells expressing podoplanin in mouse major salivary glands by laser-scanning microscopy. The expression of endothelial cell marker PECAM-1 was tested to discriminate lymphatic endothelium from salivary gland cells, and myoepithelial cells were identified by an antibody for P-cadherin. RESULTS: The podoplanin expression was rarely found in acini of the parotid gland but clearly found at the basal portion of acini in the submandibular and sublingual glands. The number of portion reacted with anti-podoplanin is greater in the sublingual gland than in the submandibular gland. The expression was also found at the basal portion of ducts in all major salivary glands. The P-cadherin expression was rarely found in acini of the parotid gland but found in acini of the sublingual gland and on ducts in parotid and sublingual glands, corresponding to the area of podoplanin expression. CONCLUSIONS: It was suggested that the acinar and myoepithelial cells in the salivary glands have the ability to express podoplanin, and that the expression may be concerned with the mucous saliva excretion.     

Monomorphic adenomas of the major and minor salivary glands. Report of twenty-one cases and review of the literature

Monomorphic adenomas are benign salivary gland tumors that have a predilection for development in the upper lip and parotid gland. Typically, patients are older persons (mean age, 61 years), but a broad age range (32 to 87 years) has been reported in the literature. Adequate treatment consists of superficial or total parotidectomy (depending on extent and location of the tumor) for parotid lesions and excision with a limited border of normal tissue for minor gland tumors. Uniform cellularity, lack of myxoid or chondroid features, and a tendency for multicentric origin are features which separate these tumors from pleomorphic adenomas. Monomorphic adenomas have been mistakenly diagnosed and treated as adenoid cystic carcinomas. Close attention to cytologic detail, histomorphology, and growth pattern at the periphery are important in separating these tumors.     

Giant pleomorphic adenoma of the parotid gland

Pleomorphic adenoma is the most common type of all benign and malignant salivary gland tumors, involving more frequently the parotid gland. It is a benign tumor with a slow and continuous growth that without treatment can reach an enormous size. We present a case of a giant pleomorphic adenoma in a 78-year-old man with a history of more than 30 years of a growing lesion in the parotid gland. Clinical examination revealed a giant mass on the right side of the face, however without any sign of facial nerve damage. The tumor was completely resected by total parotidectomy and preservation of the facial nerve. Macroscopically, the tumor measured 28 cm and weighed 4.0 Kg. On the histological examination there was a predominance of epithelial and myoepithelial cells in a hyaline and myxoid stroma. It was not found any area of malignant transformation. In the post-operatory the aesthetic and functional results were excellent.     

Metastasizing “benign” pleomorphic salivary adenoma: A dramatic case-report and literature review

Pleomorphic salivary adenomas are the most common benign neoplasms affecting the salivary glands. Very occasionally however, metastatic lesions are identified in patients with a history of PSA, which, on detailed pathological evaluation, are found to exhibit all the histological hallmarks of the preceding benign lesions.Diagnosis of benign metastasizing pleomorphic adenoma of the salivary gland is extremely rare and still under debate.We present the first case-report in literature of multiple metachronous nasal cavity, scalp and encephalic metastases of a pleomorphic adenoma of the parotid gland in a young girl.