Solid Adenoid Cystic Carcinoma Research Articles

Analysis of B7-H4 Expression Across Salivary Gland Carcinomas Reveals Adenoid Cystic Carcinoma–Specific Prognostic Relevance

B7-H4 (VTCN1), a member of the B7 family, is overexpressed in several types of cancer. Here we investigated the pattern of expression of B7-H4 in salivary gland carcinomas (SGC) and assessed its potential as a prognostic marker and therapeutic target. Immunohistochemistry (IHC) analyses were performed in a cohort of 340 patient tumors, composed of 124 adenoid cystic carcinomas (ACC), 107 salivary duct carcinomas (SDC), 64 acinic cell carcinomas, 36 mucoepidermoid carcinomas (MEC), 9 secretory carcinomas (SC), as well as 20 normal salivary glands (controls). B7-H4 expression was scored and categorized into negative (<5% expression of any intensity), low (5%–70% expression of any intensity or >70% with weak intensity), or high (>70% moderate or strong diffuse intensity). The associations between B7-H4 expression and clinicopathologic characteristics, as well as overall survival, were assessed. Among all tumors, B7-H4 expression was more prevalent in ACC (94%) compared with those of SC (67%), MEC (44%), SDC (32%), and acinic cell carcinomas (0%). Normal salivary gland tissue did not express B7-H4. High expression of B7-H4 was found exclusively in ACC (27%), SDC (11%), and MEC (8%). In SDC, B7-H4 expression was associated with female gender (P = .002) and lack of androgen receptor expression (P = .012). In ACC, B7-H4 expression was significantly associated with solid histology (P < .0001) and minor salivary gland primary (P = .02). High B7-H4 expression was associated with a poorer prognosis in ACC, regardless of clinical stage and histologic subtype. B7-H4 expression was not prognostic in the non-ACC SGC evaluated. Our comparative study revealed distinct patterns of B7-H4 expression according to SGC histology, which has potential therapeutic implications. B7-H4 expression was particularly high in solid ACC and was an independent prognostic marker in this disease but not in the other SGC assessed.

Computed Tomography and Magnetic Resonance Imaging Findings Contribute to Differentiating Solid- and Nonsolid-Type Adenoid Cystic Carcinoma in Maxillary Sinus.

This study aimed to evaluate the imaging features of maxillary sinus adenoid cystic carcinoma (ACC) on computed tomography (CT) and magnetic resonance imaging (MRI) and to investigate the imaging differences between solid and nonsolid maxillary sinus ACC. We retrospectively reviewed 40 cases of histopathologically confirmed ACC of the maxillary sinus. All the patients underwent CT and MRI. Based on the histopathological characteristics, the patients were classified into 2 groups: ( a ) solid maxillary sinus ACC (n = 16) and ( b ) nonsolid maxillary sinus ACC (n = 24). Imaging features such as tumor size, morphology, internal structure, margin, type of bone destruction, signal intensity, enhancement changes, and perineural tumor spread on CT and MRI, were evaluated. The apparent diffusion coefficient (ADC) was measured. Comparisons of imaging features and ADC values were performed between the solid and nonsolid maxillary sinus ACC using χ 2 and nonparametric tests. The internal structure, margin, type of bone destruction, and degree of enhancement significantly differed between solid and nonsolid maxillary sinus ACC (all P < 0.05). The ADC of the solid maxillary sinus ACC was considerably lower than that of the nonsolid maxillary sinus ( P < 0.05). Computed tomography and MRI may aid in the differentiation of solid and nonsolid types of maxillary sinus ACC.

Genetic heterogeneity and therapeutic target detection through microdissection in solid-type adenoid cystic carcinoma

Solid-type adenoid cystic carcinomas (ACCs) are highly aggressive and heterogeneous tumours. Because of their rarity, therapeutic strategies guided by genetic profiles based on next generation sequencing (NGS) have not been published for these tumours. Forty-nine solid-type ACCs including 43 tumours with a predominantly solid pattern, and six tumours comprising a roughly equal mixture of cribriform/tubular and solid histological forms were included in our study. The solid components from the 49 solid ACCs were enriched for mutations of genes in the NOTCH pathway (NOTCH1 61%, SPEN 24%) and chromatin remodelling pathway and the absence of myoepithelial cell differentiation. Cases with NOTCH1 mutations exhibited strong NICD expression, which was associated with poor overall and distant metastasis free survival. BRCA2 mutation and BCOR/BCORL1 mutations were observed in 20% and 18.4% of solid ACCs, respectively. In six of the solid ACCs, intratumour heterogeneity was delineated between the cribriform/tubular and solid components. NOTCH1 and FGFR2 mutations as well as NOTCH2 amplification were restricted to the solid component, indicating clonal selection within the same tumour. In two recurrent/metastatic solid ACCs, the subclones evolved in progression for local relapse and distant metastasis, although they manifested close genomic resemblance to primary tumours. Guided by the genetic profiles, the preclinical efficiency of the gamma-secretase inhibitor BMS-906024 was evaluated in patient derived xenograft models (PDXs) with activating NOTCH1 mutations and demonstrated robust antitumour effects. Our report revealed intratumour heterogeneity among solid-types within an ACC as well as the inter-tumour evolution of dominant clones among two primary and recurrent/metastatic tumours. In contrast to cribriform/tubular ACCs, solid-type ACCs should be approached with a distinct therapeutic strategy, particularly targeting NOTCH1. Microdissecting the highest grade component guided by histology is a highly recommended tumour sampling strategy and facilitates the detection of key molecular targets.

Adenoid Kistik Karsinom: Vaka Raporu

Adenoid cystic carcinoma (ACC) is the second most common malignant epithelial tumor of the salivary glands.The aim of this case report was to present a solid ACC case seen in the parotid gland. A 69-year-old male patient was admitted to our clinic with complaints of swelling, pain and paraesthesia in the posterior right mandible. In clinical examination mandibular ulcerated areas were observed. Submandibular and cervical lymphadenopathies and an expansive mass were detected in the buccal mucosa. In panoramic radiography, a radiolucent lesion was observed in the right ramus mandible, and in cone beam computed tomography(CBCT) it was determined that lesion caused bone destruction. As result of the biopsy, it was reported as ACC and excision was recommended. Early diagnosis and treatment is very important in terms of survival and prevention of metastasis to distant areas. Therefore, accurate diagnosis and evaluation will affect the treatment success.

Notch activation leads to loss of myoepithelial differentiation and poor outcome in solid adenoid cystic carcinoma.

We aimed to investigate Notch pathway dysregulation in solid adenoid cystic carcinoma (AdCC) and to define the association of Notch activation with cell differentiation and prognosis in AdCCs. Notch1 mutations were detected from 125 AdCCs (62 cribriform-tubular; 63 solid). RNA-seq was performed in 16 AdCCs (6 Notch-mutant; 10 wild type). Notch activation indicator NICD and myoepithelial marker p63 were detected using immunohistochemistry and double-labelling immunofluorescence. The effect of exogenous NICD overexpression on p63 expression and cell proliferation was investigated using Western blotting and live-cell imaging. We identified 33 Notch1 activating mutations in 27 AdCCs including 26 solid and 1 cribriform-tubular subtypes. Six tumours harboured more than one Notch1 mutation, and 18 Notch1 mutations were novel. Most (47/63, 74.6%) solid AdCCs showed NICD overexpression, whereas 61 of 62 (98.4%) cribriform-tubular tumours were negative. NICD and p63 exhibited mutually exclusive expression, and exogenous NICD overexpression promoted cell proliferation and decreased p63 expression. NICD overexpression and Notch mutations were poor indicators for overall survival and metastasis, especially bone metastasis. Dysregulated Notch signalling plays a critical role in AdCC severity. Notch activation may contribute to loss of myoepithelial differentiation as well as high proliferation and metastasis rates in solid AdCC.

Solid-type adenoid cystic carcinoma of the breast, a distinct molecular entity enriched in NOTCH and CREBBP mutations.

Adenoid cystic carcinoma (ACC) of the breast with a predominant solid pattern is difficult to diagnose with certainty and differentiate from more common triple-negative breast cancers (TNBCs) of basal-phenotype. To better characterize solid ACC, we performed a clinical, morphological, immunohistochemical, and molecular comparative analysis of 33 ACCs of the breast comprising 17 solid variant ACCs and 16 conventional ACCs. Solid ACCs displayed basaloid morphology with an exclusive or predominant epithelial cell population associated with decreased myoepithelial differentiation, while demonstrating MYB protein overexpression similar to the more common type of ACC. Strong and diffuse MYB expression by immunochemistry was observed in 14/17 (82%) of solid ACCs while MYB rearrangements were detected by break apart fluorescence in situ hybridization (FISH) in only 3/16 (19%) of solid ACCs. Conversely, weak MYB immunohistochemical expression was observed in only 7/204 (3%) of TNBC. Solid ACCs displayed a transcriptomic profile distinct from conventional ACCs with 549 genes showing a highly significant differential expression between conventional and solid ACC [false discovery rate (FDR) < 0.01; log2FC > |1|]. EnrichR and Kegg Pathway analyses identified PI3K-Akt and focal adhesion signaling pathways as significantly overexpressed in conventional ACCs compared with solid ACCs which significantly overexpressed the nitrogen metabolism pathway. CREBBP mutations and NOTCH activating gene mutations were only present in solid ACCs, concerning 5/16 (31%) of cases for each gene. Tumors with NOTCH activating mutations displayed a strong diffuse nuclear NICD1 staining, an established marker of Notch pathway activation. Solid ACCs also differed from basal-type TNBC, with fewer TP53 mutations and a more stable genomic profile on array comparative genomic hybridization (CGH). In summary, solid-type ACC of the breast is a distinct molecular entity within the ACC family and is different from common basal-type TNBC. MYB is a diagnostically useful biomarker of solid ACC and NOTCH could be a novel potential therapeutic target in 30% of cases.

Adenoid cystic carcinoma: immunohistochemistry and differential diagnosis, a case report

ABSTRACT A 52-year-old male patient complained of loss of sensitivity and pain in the maxilla. After examination, root canal treatment of tooth 12 was requested. Subsequently, there was a purplish increased volume of softened consistency in the area of vestibular attached gingiva of tooth 12. The anterior upper dental segment presented mobility. Incisional biopsy revealed malignant neoplasm of glandular epithelial origin, suggestive of solid adenoid cystic carcinoma (ACC). An immunohistochemical panel was performed, which confirmed the histopathological suspicion. Solid ACC may lead to diagnostic difficulties, since this lesion exhibits morphological features similar to other malignancies.

P120 catenin expression and its correlation with E-cadherin in salivary gland neoplasms

ObjectivesAltered P120 catenin expression has been associated with E-cadherin loss and poor prognosis in several cancers. The objectives of this study were to examine the P120 catenin expression in salivary gland neoplasms in correlation with E-cadherin and assess the relationships between their expression levels and pathologic characteristics. MethodsFifty-two cases of salivary gland neoplasms, i.e. 25 mucoepidermoid carcinomas (MEC), 13 adenoid cystic carcinomas (ACC), 12 pleomorphic adenomas (PA) and 2 polymorphous adenocarcinomas (PAC) were included. The expression of P120 catenin and E-cadherin was investigated immunohistochemically. ResultsBoth P120 catenin and E-cadherin were overexpressed in salivary gland neoplasms, compared to normal tissue. P120 catenin was primarily detected on the membrane of neoplastic cells in most cases. A significant correlation between levels of expression of both proteins was noted in MECs. In ACCs and PA, ductal cells showed positive immunoreactivity, whereas myoepithelial cells variably expressed both proteins. Increased P120 catenin expression was significantly associated with the solid subtype of ACCs. ConclusionsThe cadherin-catenin complex is preserved in the heterogenous tumor cell population in salivary gland neoplasms. Overexpression of P120 catenin may be involved in the progression to solid ACCs.

A possible correlation of Ezrin and Fascin-1 in adenoid cystic carcinoma of salivary gland: an immunohistochemical study

Adenoid cystic carcinoma (ACC) of salivary gland is one of the most common epithelial salivary gland carcinomas and is characterized by extensive local tissue infiltration and poor long-term survival. All forms of ACC are characterized by prolonged but persistent growth, extensive perineural invasion and hematogenous metastases. Distant sites of metastasis are primarily lung, although bone and liver are also common. The incremental motility of malignant cells is a critical step in their migration, invasion and metastasis, which is regulated by reorganization of actin cytoskeleton and regulation of focal adhesion. Fascin-1 and ezrin are essential components of these cellular structures. The aim of this study is to evaluate the expression of these cytoskeleton-associated proteins in ACC as a prognostic indicator of invasiveness. Five cases of normal salivary gland tissue and 43 cases of adenoid cystic carcinoma (10 cases cribriform form, 15 cases tubular form and 18 cases solid form). Immunohistochmical staining for ezrin and fascin-1 antibodies were done for all specimens. Statistical analysis was performed using a commercially available software program (SPSS 19; SPSS, Chicago, IL, USA). The level of significance was set at P < 0.05. For both markers, statistically, the greatest mean area percent was recorded in solid pattern ACC, whereas the lowest values were recorded in normal tissue. These results indicating a synergistic effect between ezrin and fascin-1 during ACC invasion.

Molecular features of adenoid cystic carcinoma with an emphasis on microRNA expression.

Molecular features of adenoid cystic carcinoma with an emphasis on microRNA expression.

Assessment of CTNNB1 gene mutations and β-catenin immunoexpression in salivary gland pleomorphic adenomas and adenoid cystic carcinomas.

β-Catenin exerts multiple functions in several neoplasms, playing a major role in cell signaling and tumor progression. This study analyzed possible CTNNB1 mutations in salivary gland pleomorphic adenomas (PAs) and adenoid cystic carcinomas (ACCs), and determined possible differences in β-catenin immunoexpression in relation to these mutations, as well as histopathological aspects of these tumors. Twenty-four PAs (15 cell-rich and 9 cell-poor tumors) and 24 ACCs (10 tubular, 8 cribriform, and 6 solid tumors) were selected for the analysis of β-catenin distribution and cellular localization. Furthermore, β-catenin expression was evaluated using the H-score scoring system. Mutations in CTNNB1 exon 3 were investigated by the single-strand conformational polymorphism test. Diffuse β-catenin expression was more frequently observed in ACCs compared to PAs (P = 0.008). No significant difference in β-catenin cellular localization was observed between these tumors (P = 0.098). Comparisons between PA and ACC cases revealed a higher median H-score in the latter (P = 0.036). Cell-rich PAs exhibited a trend for higher H-score than cell-poor tumors (P = 0.060), whereas lower H-scores were observed in cribriform ACCs when compared to tubular and solid ACCs (P = 0.042). Mutations in CTNNB1 were observed in 6 PAs and 7 ACCs, with no significant difference in H-scores for β-catenin according to mutation status (P = 0.135). β-Catenin is important in the pathogenesis of salivary gland PAs and ACCs. In addition, CTNNB1 exon 3 mutations do not seem to significantly influence β-catenin cytoplasmic/membranous expression or nuclear translocation in these tumors.

Usefulness of NKX2.2 Immunohistochemistry for Distinguishing Ewing Sarcoma from Other Sinonasal Small Round Blue Cell Tumors.

NKX2.2 is a new immunohistochemical marker that has been reported to be sensitive and specific for Ewing sarcoma (ES). It has not, however, been investigated specifically in the sinonasal small round blue cell tumor (SRBCT) differential diagnosis which includes many tumors specific to that site. It has also not been investigated in the newly recognized "adamantinoma-like" variant of ES. Immunohistochemistry for NKX2.2 was performed on 170 poorly differentiated sinonasal neoplasms: 73 squamous cell carcinomas (67 poorly differentiated, non-keratinizing, or basaloid types and 6 nasopharyngeal carcinomas), 46 olfactory neuroblastomas, 8 sinonasal undifferentiated carcinomas (SNUCs), 6 melanomas, 7 Ewing sarcomas, 6 SMARCB1-deficient carcinomas, 6 teratocarcinosarcomas, 5 alveolar rhabdomyosarcomas, 4 solid adenoid cystic carcinomas, 4 NK/T cell lymphomas, 3 NUT carcinomas, and 2 small cell carcinomas. NKX2.2 was positive in 7 of 7 (100%) Ewing sarcomas, including 3 adamantinoma-like variant (all diffuse, 5 strong and 2 weak). It was also positive in 5 of 6 (83%) teratocarcinosarcomas (strong, but focal), 12 of 46 (26%) olfactory neuroblastomas (diffuse, 2 strong and 10 weak), 4 of 6 melanomas (2 diffuse, 2 focal, all weak), and 1 of 2 small cell carcinomas (diffuse and strong). All squamous cell carcinomas, NUT carcinomas, SMARCB1-deficient carcinomas, SNUCs, solid adenoid cystic carcinomas, NK/T cell lymphomas, and alveolar rhabdomyosarcomas were negative. In the sinonasal SRBCT differential diagnosis, NKX2.2 is a useful and very sensitive marker for Ewing sarcoma, including the treacherous adamantinoma-like variant. At the same time, it is not entirely specific, as it will be positive in a subset of other neuroendocrine/neuroectodermal tumors. As a result, NKX2.2 must be utilized as part of an immunohistochemical panel with other markers, especially cytokeratins, melanoma markers, and CD99.

Genetic events in the progression of adenoid cystic carcinoma of the breast to high-grade triple-negative breast cancer

AbstractAdenoid cystic carcinoma of the breast is a rare histological type of triple-negative breast cancer with an indolent clinical behavior, often driven by the MYB-NFIB fusion gene. Here we sought to define the repertoire of somatic genetic alterations in two adenoid cystic carcinomas associated with high-grade triple-negative breast cancer. The different components of each case were subjected to copy number profiling and massively parallel sequencing targeting all exons and selected regulatory and intronic regions of 488 genes. Reverse transcription PCR and fluorescence in situ hybridization were employed to investigate the presence of the MYB-NFIB translocation. The MYB-NFIB fusion gene was detected in both adenoid cystic carcinomas and their associated high-grade triple-negative breast cancer components. Although the distinct components of both cases displayed similar patterns of gene copy number alterations, massively parallel sequencing analysis revealed intratumor genetic heterogeneity. In case 1, progression from the trabecular adenoid cystic carcinoma to the high-grade triple-negative breast cancer was found to involve clonal shifts with enrichment of mutations affecting EP300, NOTCH1, ERBB2 and FGFR1 in the high-grade triple-negative breast cancer. In case 2, a clonal KMT2C mutation was present in the cribriform adenoid cystic carcinoma, solid adenoid cystic carcinoma and high-grade triple-negative breast cancer components, whereas a mutation affecting MYB was present only in the solid and high-grade triple-negative breast cancer areas and additional three mutations targeting STAG2, KDM6A and CDK12 were restricted to the high-grade triple-negative breast cancer. In conclusion, adenoid cystic carcinomas of the breast with high-grade transformation are underpinned by the MYB-NFIB fusion gene and, akin to other forms of cancer, may be constituted by a mosaic of cancer cell clones at diagnosis. The progression from adenoid cystic carcinoma to high-grade triple-negative breast cancer of no special type may involve the selection of neoplastic clones and/or the acquisition of additional genetic alterations.

Characterization and localization of c-kit and epidermal growth factor receptor in different patterns of adenoid cystic carcinoma.

Adenoid cystic carcinoma (ACC) is malignant neoplasm of the salivary glands. It accounts for most cases of minor salivary gland malignancies and a substantial proportion of parotid and submandibular gland malignancies. ACC is associated with a high mortality rate, and it often recurs after prolonged periods of time, and this occurs even when radical excision has been performed. The present study was aimed to determine the localization of dual cell population and to analyze the potency of using a system of dual markers (c-kit and epidermal growth factor receptor [EGFR]) in enhancing the characterization of ACC. Three micrometer thin sections of adenoid cystic carcinoma were obtained. One set of slides was stained by hematoxylin and eosin for reconfirmation of histological diagnosis while the other two sets were stained for c-kit and EGFR using immunohistochemical method. Statistical Analysis Used and Results: Show c-kit expression to be limited to the inner ductal epithelial cells and the EGFR expression mainly to the outer myoepithelial cells in the majority of tubular and cribriform patterns. In solid ACC, c-kit was uniformly positive while EGFR was consistently negative. C-kit and EGFR biomarkers can be used to enhance the characterization of ACC and to determine the localization of dual cell population which could suggest the dual origin of ACC and provides evidence for the new therapeutic strategy in ACC.

High frequency of loss of PTEN expression in human solid salivary adenoid cystic carcinoma and its implication for targeted therapy.

Salivary gland tumor (SGT) is one of the least studied cancers due to its rarity and heterogeneous histological types. Here, we reported that loss of PTEN expression was most frequently found in the poorly differentiated, high grade solid adenoid cystic carcinomas. Loss of PTEN expression correlated with activation of mTOR by increased phosphorylated S6 ribosome protein. We further functionally studied the role of PTEN in a pair of human SACC cell lines, SACC-83 and SACC-LM. Reduced PTEN level was correlated with the metastasis potential. When we knocked down PTEN in the SACC-83 cell line, we observed increased proliferation and enhanced migration/invasion in vitro, and increased tumor size in vivo. We further tested the therapeutical effect by applying a PI3K/mTOR inhibitor NVP-BEZ235 to both SACC cell lines. Decreased cell proliferation, increased apoptosis, as well as reduced cell migration/invasion were observed in both cell lines upon the NVP-BEZ235 treatment. Moreover, the NVP-BEZ235 treatment in a SGT xenograft mouse model significantly reduced primary tumor size and lung metastasis. Taken together, our results demonstrated that PTEN is a potent tumor suppressor in human SGTs, and targeting PI3K/mTOR pathway may be effective in the targeted therapy for human SGT patients with loss of PTEN expression.

Histopathological grading of adenoid cystic carcinoma of the head and neck: Analysis of currently used grading systems and proposal for a simplified grading scheme

Histopathological grading of adenoid cystic carcinoma (ACC) is a controversial issue. It is generally agreed that solid type ACC has a relatively poor prognosis. However, the amount of solid regions within this often mixed type tumor that predicts a poor prognosis is not firmly established. Some authors stipulate that the presence of a solid component regardless of the amount is a poor prognosticator where others argue that the amount should be taken into consideration. Two grading systems most commonly used are those described by Perzin et al./Szanto et al. and Spiro et al., respectively. They report that prognosis of ACC is poor if >30% and >50% of the tumor volume has a solid growth pattern, respectively. The described grading systems are applied to a series of 81 surgically treated cases of ACC at the VU University Medical Center, Amsterdam, The Netherlands. Moreover, we introduced an alternative grading system, in which the presence of a solid component, irrespective of its amount, is considered. All three systems of grading were tested for inter-observer concordance and prediction of prognosis. Inter-observer concordance for grading ACC according to Perzin et al./Szanto et al. and Spiro et al., proved to be moderate with Kappa Scores of 0.393 and 0.433, respectively. Our alternative grading system yielded inter-observer concordance with a Cohen's kappa result of 0.990. All systems were comparable in discriminating patients with poor clinical outcome. Histopathological grade proved to be an independent prognosticator. The presence of any solid component in ACC is a negative prognosticator, and can histopathologically be diagnosed with a high reliability. These results suggest to merely register the presence or absence of a solid tumor component since its inter-observer variability is very low, its reproducibility is high and its predictive value is comparable to the traditional grading systems used.

Utility of p40 in the Differential Diagnosis of Small Round Blue Cell Tumors of the Sinonasal Tract

The sinonasal tract may give rise to a broad range of neoplasms that share a "small round blue cell" tumor (SBRCT) appearance on routine histology, but treatment strategies depend on precise tumor classification. Immunohistochemistry for p63 is often employed in the sinonasal SRBCT differential diagnosis because it is highly sensitive for squamous cell carcinoma (SCC). However, p63 staining may be observed in other tumor types, a potential diagnostic pitfall. P40 is a more squamous-specific isoform of p63, and it may be more useful in distinguishing poorly differentiated SCC from its mimickers in the sinonasal tract. Immunohistochemistry for p40 and p63 was performed on 171 sinonasal neoplasms with SRBCT morphology: 73 SCCs (67 poorly differentiated, non-keratinizing, or basaloid types and 6 nasopharyngeal carcinomas), 46 esthesioneuroblastomas, 11 sinonasal undifferentiated carcinomas (SNUCs), 11 lymphomas, 9 melanomas, 7 alveolar rhabdomyosarcomas, 4 solid adenoid cystic carcinomas, 4 NUT midline carcinomas, 4 primitive neuroectodermal tumors (PNETs), and 2 small cell carcinomas. P40 was positive in 72 of 73 SCCs, and showed a diffuse distribution in all but one positive case. P40 immunoexpression was also observed in 13 of 46 (28%) esthesioneuroblastomas, 6 of 11 (55%) SNUCs, 2 of 4 (50%) adenoid cystic carcinomas, 3 of 4 (75%) NUT midline carcinomas, 1 of 2 (50%) small cell carcinomas, and 1 of 4 (25%) PNETs; in the non-SCC tumors, p40 staining was focal in most cases. P63 was positive in every p40-positive tumor. In addition, a p63+/p40- phenotype was seen 5 of 11 (45%) lymphomas, 4 of 7 (57%) alveolar rhabdomyosarcomas, 1 of 4 (25%) PNETs, and 3 of 46 (7%) esthesioneuroblastomas. All sinonasal melanomas were negative for both markers. In the sinonasal SRBCT differential diagnosis, both p40 and p63 are highly sensitive for SCC, but p40 is more specific. Notably, p40 is consistently negative in lymphomas and alveolar rhabdomyosarcomas, two tumors that are frequently p63-positive. It must be remembered, however, that even diffuse p40 immunostaining is not entirely specific for the squamous phenotype, and therefore it should be utilized as part of an immunohistochemical panel.

Immunoexpression of α2β1, α3β1, and α5β1 Integrins in Pleomorphic Adenoma and Adenoid Cystic Carcinoma

The aim of the present study was to compare the expression of α2β1, α3β1, and α5β1 integrins between 28 pleomorphic adenomas (PAs) and 10 adenoid cystic carcinomas (ACCs), and investigate differences in the expression of these integrins according to histologic subtypes of ACCs. It was taken into consideration the presence or absence, distribution, and localization of integrin immunoexpression. There was immunoreactivity in the intercellular contacts of the strands, nests, and solid sheets of PAs, as well as in the luminal and nonluminal cells of the duct-like structures, with a predominant immunoexpression in the luminal cells. The immunoexpression in ACCs varied with histologic subtype of the tumor. It was verified for a tendency of absence and/or reduced expression of all integrins in the solid subtype of ACCs. In general, PAs revealed a more diffuse and remarkable immunoexpression of all studied integrins than ACCs. The reduced integrins expression in ACC may be related to a lesser degree of cell differentiation in this neoplasm. Moreover, the absence and/or reduced expression of the studied integrins in solid ACC suggest a possible role in pathogenesis and more aggressive biological behavior of this histologic subtype.

Developmental transcription factor EN1—a novel biomarker in human salivary gland adenoid cystic carcinoma*

Adenoid cystic carcinoma (ACC), a rare and progressive salivary malignancy, is characterized by histogenetic, morphologic, and clinical heterogeneity. Extensive efforts to characterize the molecular events associated with these tumors have included the identification of biomarkers for prognostication and post-therapy assessment. In a previous study of genome-wide methylation screening, the authors of the current report identified a limited number of differentially methylated gene regions in ACC, and significant hypermethylation was observed at the transcriptional start sites of genes that encode for the transcription factor engrailed homeobox 1 (EN1). Clinicopathologic correlation analyses indicated that EN1 methylation status is correlated with histologic tumor grade, tumor location, and final patient outcome. To ascertain definitively whether aberrant EN1 expression accompanies human salivary ACC, the authors used an immunohistochemical technique to directly evaluate EN1 protein expression in ACC of the salivary gland. The data revealed increased EN1 protein expression in solid type ACC, which was correlated with a significantly lower survival rate. The current results validated EN1 as a potential biomarker in a large cohort of patients with salivary ACC. Immunohistochemical analysis of EN1 in biopsy specimens obtained for diagnostic purposes and/or surgically resected material may reveal that EN1 is a biologic predictor of poor prognosis in patients with salivary ACC.

Genetic profile of adenoid cystic carcinomas (ACC) with high-grade transformation versus solid type

ACC can occasionally undergo dedifferentiation also referred to as high-grade transformation (ACC-HGT). However, ACC-HGT can also undergo transformation to adenocarcinomas which are not poorly differentiated. ACC-HGT is generally considered to be an aggressive variant of ACC, even more than solid ACC. This study was aimed to describe the genetic changes of ACC-HGT in relation to clinico-pathological features, and to compare results to solid ACC. Genome wide DNA copy number changes were analyzed by microarray CGH in ACC-HGT, four with transformation into moderately differentiated adenocarcinoma (MDA) and two into poorly differentiated carcinoma (PDC), and five solid ACC. In addition, Ki67 index and p53 immunopositivity was assessed. ACC-HGT carried fewer copy number changes compared to solid ACC. Two ACC-HGT cases harboured a breakpoint at 6q23, near the cMYB oncogene. The complexity of the genomic profile concurred with the clinical course of the patient. Among the ACC-HGT, p53 positivity significantly increased from the conventional to the transformed (both MDA and PDC) component. ACC-HGT may not necessarily reflect a more advanced stage of tumor progression, but rather a transformation to another histological form in which the poorly differentiated forms (PDC) presents a genetic complexity similar to the solid ACC.