SET protein accumulation prevents cell death in head and neck squamous cell carcinoma through regulation of redox state and autophagy.

Molecular alterations in cell death pathways and imbalances in regulators of up- or downstream signaling pathways can lead to resistance to cell death, which is one of the hallmarks of cancer. These signaling modifications are strategies that tumor cells use to resist chemotherapy and that contribute to the high recurrence rate of head and neck squamous cell carcinoma (HNSCC). The SET oncoprotein is a PP2A inhibitor that accumulates in HNSCC and represents a promising therapeutic target. Here we report the role that SET protein plays in resistance to death of two HNSCC cell lines: Cal 27 and HN13. SET protein regulated intracellular redox balance by controlling cellular localization of APE 1 - an endonuclease that is part of the SET complex and regulates antioxidant gene transcription. SET protein knockdown (siSET) associated with tert-butyl hydroperoxide-induced oxidative stress sensitized Cal 27 and HN13 cells to apoptosis via the extrinsic and intrinsic pathways, respectively. SET protein upregulated autophagy in HNSCC cells in a PP2A-dependent manner and apparently regulated ULK1 expression. The fact that siSET lowered Bcl-2 phosphorylation levels indicated that SET protein interfered with an alternative pathway that modulated autophagy in HNSCC cells. Overall, SET protein regulated intracellular redox state and sustained autophagy in HNSCC cells, which may explain resistance to death of HNSCC cells. Altogether, the findings reported herein support SET protein as therapeutic target for HNSCC.

Cervical ganglioneuroblastoma in a new born Japanese Black calf.

This case report describes a congenital ganglioneuroblastoma in a 38-day-old male Japanese Black calf. The cervical multinodular mass was present at birth and grew rapidly. The cut surface was pale gray-to-yellow and had a gelatinous appearance. Hemorrhagic cysts of various sizes were observed in the nodule. Histologically, the mass contained clusters of neuroblastic cells, ganglionic cells, and Schwann-like cells. Immunohistochemically, the ganglionic cells showed strong positivity for neuron-specific enolase, neurofilament, synaptophysin, and chromogranin A, whereas the Schwann-like cells strongly expressed S-100, glial fibrillary acidic protein, and vimentin. Ultrastructurally, neurosecretory granules resembling catecholamine were observed in the neuroblastic and Schwann-like cells. Based on the pathology, the diagnosis was congenital cervical nodular ganglioneuroblastoma.

Does pTis exist in HPV-driven tonsillar carcinomas? An ultrastructural review and examination of two cases.

Many tonsillar tumors present clinically as cervical nodal metastases and the primary tumor is often difficult to find. HPV-driven tonsillar carcinoma begins in the reticulated crypt epithelium, possibly through viral integration. The basement membrane is not complete in the reticulated crypt epithelium, which may enhance the immune function. We examined the reticulated crypt epithelium in a normal case and five neoplastic tonsils with cervical metastasis as the presenting symptom to further investigate whether tonsil carcinoma in-situ exists. Our results suggest that in-situ carcinoma may need to be excluded from the future staging for human papilloma virus associated tonsillar tumors.

Head and Neck Extranodal Interdigitating Dendritic Cell Sarcoma: Case Report and Review of the Literature.

Interdigitating dendritic cell sarcoma (IDCS) is an exceedingly rare neoplasm originating from professional antigen presenting cells normally located in the T zone of the lymph node. The purpose of this report was to describe the first case of the IDCS of the submandibular gland and perform a review of the literature of head and neck IDCS. We present a case of an 81-year-old man with a 5 months history of slowly enlarging painless mass in right submandibular region. Fine needle aspiration cytology was suggestive of squamous cell carcinoma. The patient underwent surgical resection of the right submandibular gland and neck dissection. A malignant spindle cell proliferation involving the submandibular gland and colonizing one laterocervical lymph node was found. Morphology and immunophenotype prompted a differential diagnosis of a metastatic spindle cell melanoma versus an IDCS. Transmission electron microscopy was performed and supported a diagnosis of IDCS. The diagnosis of IDCS is a challenging task and may require a large array of techniques.

Nuclear localization of MUC1 extracellular domain in breast, head and neck, and colon cancer.

BACKGROUND: The glycoprotein MUC1 is overexpressed and underglycosylated in cancer cells. MUC1 is translated as a single polypeptide that undergoes autocleavage into 2 subunits (the extracellular domain and the cytoplasmic tail), and forms a stable heterodimer at the apical membrane of normal epithelial cells. The MUC1 cytoplasmic tail localizes to the cytoplasm of transformed cells and is targeted to the nucleus. AIMS: To study the expression of the MUC1 extracellular subunit in cell nuclei of neoplastic breast, head and neck, and colon samples. MATERIALS AND METHODS: 330 primary tumor samples were analyzed: 166 invasive breast carcinomas, 127 head and neck tumors, and 47 colon tumors; 10 benign breast disease (BBD) and 40 normal specimens were also included. A standard immunohistochemical method with antigen retrieval was performed. Nuclear fractions from tissue homogenates and breast cancer cell lines (ZR-75, MDA-MB-231, MCF7, and T47D) were obtained and analyzed by Western blotting (WB). The anti-MUC1 extracellular subunit monoclonal antibody HMFG1 was used for immunohistochemistry. RESULTS: 37/166 breast cancer specimens, 5/127 head and neck cancer specimens, 2/47 colon cancer samples, and 3/10 BBD samples showed immunohistochemical staining at the nuclear level. No nuclear reaction was detected in normal samples. By WB, breast and colon cancer purified nuclear fractions showed reactivity at 200 kDa in 3/30 breast and 3/20 colon cancer samples as well as purified nuclear fractions obtained from breast cancer cell lines. CONCLUSIONS: This study shows that the MUC1 extracellular domain might be translocated to the cell nucleus in breast, head and neck, and colon cancer as well as BBD.

Primary ductal adenocarcinoma of the lacrimal gland, associated with abundant intracytoplasmic lumens containing some eosinophilic hyaline globules: cytological, histological and ultrastructural findings.

A primary ductal adenocarcinoma (PDA) of the lacrimal gland is a rare distinct subtype of an epithelial tumor arising in the lacrimal gland. PDA is the counterpart of salivary duct carcinoma (SDC) resembling an invasive ductal carcinoma (IDC) of the breast. In our case, PDA revealed histopathological and immunohistochemical results corresponding to SDC. Interestingly, the tumor cells showed intracytoplasmic vacuoles containing dense eosinophilic hyaline globules at light microscopy. Ultrastructurally, the tumor cells exhibited microvilli-lined intracytoplasmic lumen containing homogenous electron-dense secretory products. A previous study demonstrated that numerous intracytoplasmic lumens of tumor cells are favored breast malignant tumor, similar to the histopathology of PDA, rather than benign lesion. This characteristic finding may be meaningful to diagnose high grade epithelial tumors including PDA.

Identification of cancer stem-like side population cells in purified primary cultured human laryngeal squamous cell carcinoma epithelia.

Cancer stem-like side population (SP) cells have been identified in many solid tumors; however, most of these investigations are performed using established cancer cell lines. Cancer cells in tumor tissue containing fibroblasts and many other types of cells are much more complex than any cancer cell line. Although SP cells were identified in the laryngeal squamous cell carcinoma (LSCC) cell line Hep-2 in our pilot study, it is unknown whether the LSCC tissue contains SP cells. In this study, LSCC cells (LSCCs) were primary cultured and purified from a surgically resected LSCC specimen derived from a well-differentiated epiglottic neoplasm of a Chinese male. This was followed by the verification of epithelium-specific characteristics, such as ultrastructure and biomarkers. A distinct SP subpopulation (4.45±1.07%) was isolated by Hoechst 33342 efflux analysis from cultured LSCCs by using a flow cytometer. Cancer stem cell (CSC)-associated assays, including expression of self-renewal and CSC marker genes, proliferation, differentiation, spheroid formation, chemotherapy resistance, and tumorigenicity were then conducted between SP and non-SP (NSP) LSCCs. In vitro and in vivo assays revealed that SP cells manifested preferential expression of self-renewal and CSC marker genes, higher capacity for proliferation, differentiation, and spheroid formation; enhanced resistance to chemotherapy; and greater xenograft tumorigenicity in immunodeficient mice compared with NSP cells. These findings suggest that the primary cultured and purified LSCCs contain cancer stem-like SP cells, which may serve as a valuable model for CSC research in LSCC.

Neuroendocrine differentiation in head and neck squamous cell carcinoma.

OBJECTIVE: Tumours with neuroendocrine differentiation frequently express chromogranin A, synaptophysin and somatostatin receptors. The role of neuroendocrine differentiation in head and neck squamous cell carcinoma is not yet clear. METHOD: The presence of chromogranin A, synaptophysin and somatostatin receptors was studied immunohistochemically in 78 head and neck squamous cell carcinoma specimens. RESULTS: Sparse chromogranin A expression was found in 41 per cent, associated with high chromogranin A messenger RNA expression and the presence of dense core granules. Low synaptophysin expression was found in 18 per cent. The highest staining scores were found for somatostatin receptor 5 (82 per cent), followed by somatostatin receptor 1 (69 per cent) and somatostatin receptor 2 (54 per cent), whereas somatostatin receptors 3 and 4 expression was low. Expression was not correlated with tumour stage or survival. CONCLUSION: Cells with neuroendocrine differentiation are sparsely scattered in some head and neck squamous cell carcinomas. Their pathophysiological role is elusive. In contrast, somatostatin receptor and particularly somatostatin receptor 5 expression is frequent in head and neck squamous cell carcinoma. Somatostatin receptor expression is not considered to indicate neuroendocrine differentiation in head and neck squamous cell carcinoma.

Oral and oropharyngeal squamous cell carcinoma in our population: the clinic-pathological and morphological description of 153 cases / Carcinoma de células escamosas oral y orofaríngeo en nuestra población: descripción clínico-patológica y morfológica de 153 casos

Oral cavity and oropharynx are one of the commonest sites for cancers in our population due to a high prevalence of tobacco use, especially chewing, in our country. The objective of this study was to prospectively assess the clinico-pathological and morphological aspects of the patients presenting to us with these malignancies. We studied 153 patients with oral and oropharyngeal squamous cell carcinoma who were managed in the department of otolaryngology and head and neck surgery between January 2006 and December 2007 at Maulana Azad Medical College and associated hospitals in New Delhi. There were 127 male patients (83 percent) and 26 females (17 percent) with ages ranging from 22 years to 70 years. One hundred and eleven patients (73 percent) presented with oral cavity and 42 patients (27 percent) with oropharyngeal carcinomas. Most common site of presentation in the oral cavity was the buccal mucosa, whereas, base of tongue was the commonest site in the oropharynx. Amongst the oral cavity cancers, 51 each (46 percent each) were well differentiated and moderately differentiated, whereas, 9 (8 percent) were poorly differentiated cancers. However, amongst the oropharyngeal cancers, 27 (64 percent) were moderately differentiated and 15 (36 percent) were poorly differentiated. Overall, 73 patients presented with lymphadenopathy on presentation, out of which, 44 patients were those with oral cavity and 29 with oropharyngeal cancers. There was a significant correlation between the site (i.e. oral cavity or orophaynx) and lymphadenopathy on presentation. Fifty nine patients (39 percent) presented to us with early stage disease (i.e. stage I and II), whereas, 94 patients (61 percent) presented with late stage disease (i.e. stage III and IV). There was a significant correlation between the site (i.e. oral cavity or oropharynx) and stage at presentation.

En nuestro país, la cavidad oral y orofaringe son los sitios más comunmente afectados por cáncer, debido a la alta prevalencia del consumo de tabaco, especialmente, el tabaco de mascar. El objetivo de este estudio fue evaluar prospectivamente los aspectos clínico-patológicos y morfológicos de los pacientes que acuden con estas malignidades. Se estudiaron 153 pacientes con carcinoma de células escamosas de la cavidad oral y orofaringe que fueron tratados en el Servicio de Otorrinolaringología y Cirugía de Cabeza y Cuello entre enero del 2006 y diciembre del 2007 en Maulana Azad Medical College y hospitales asociados en Nueva Delhi. Se evaluaron pacientes de ambos sexos, 127 hombres (83 por ciento) y 26 mujeres (17 por ciento) con edades entre 22 a 70 años. Ciento once pacientes (73 por ciento) presentaron carcinoma de la cavidad oral y 42 (27 por ciento) carcinomas de la orofaringe. El sitio más común de presentación en la cavidad oral fue la mucosa bucal, mientras que la base de la lengua fue el sitio más común de la orofaringe. Entre los cánceres de la cavidad oral, 51 (46 por ciento cada uno) estaban bien diferenciados y moderadamente diferenciados, mientras que 9 (8 por ciento) fueron escazamente diferenciado. Sin embargo, entre los cánceres de la orofaringe, 27 (64 por ciento) fueron moderadamente diferenciados y 15 (36 por ciento) fueron pobremente diferenciados. En total, 73 pacientes se presentaron con adenopatías, de los cuales, 44 pacientes fueron en la cavidad oral y 29 en la orofaringe. Se observó una correlación significativa entre el sitio (es decir, la cavidad oral u orofaringe) y la presentación de adenopatías. Cincuenta y nueve pacientes (39 por ciento) se presentaron con enfermedad en estadio temprano (estadios I y II), mientras que 94 pacientes (61 por ciento) se presentaron con enfermedad en estadio tardío (etapas III y IV). Se observó una correlación significativa entre el sitio (cavidad oral u orofaringe) y la etapa en la presentación.

[Morphological bases of dopamine effect on HEP-2 tumor cells viability].

The purpose of the present investigation was to study the morpho-functional organization of a classical object of cytological research - cultured HEp-2 tumor cells, using dopamine as a penetrating agent, inducing the polymerization of cytosolic actin. It was demonstrated that dopamine introduced into the incubation medium reduced viability and caused morphological disturbances of cultured HEp-2 cells; these effects were proportional to dopamine concentrations (1.0 x 10(-4) M to 1.0 x 10(-3) M) and exposure duration (2 to 3 days). These cells, according to ultrastructural data, underwent fusion and lysis because of the appearance of actin filaments network in the loci of globular actin prevalence in control cells. Dopamine receptors had no effect on cytotoxic effect of dopamine. This was indicated by fluorescent microscopical evidence of dopamine penetration into experimental cells in the presence of haloperidol, as well as destruction of HEp-2 cells under the action of pyrimidinethione, similar to dopamine by characteristics, but lacking its own receptors. It is suggested that cytoplasmic target for dopamine is globular actin and that induced polymerization of this cytoskeletal protein caused injury to tumour cells.

Zinc oxide nanoparticles induce photocatalytic cell death in human head and neck squamous cell carcinoma cell lines in vitro.

The aim of this study was to determine the photocatalytic effects of zinc oxide (ZnO) NPs in combination with UVA-1 in human head and neck squamous cell carcinoma (HNSCC) cell lines in vitro. NP characteristics and intracellular distribution were described by transmission electron microscopy (TEM). After pre-incubation with ZnO NPs in concentrations of 0.002-20 µg/ml, the HNSCC cell lines HLaC 78 and UD-SCC 7A as well as primary oral mucosa cells (pOMCs) were treated with UVA-1. Cell survival and vitality was observed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide-(MTT)-assay and fluorescein diacetate test. Apoptosis was assessed by annexin-V propidium iodide flow cytometry. Intranuclear distribution of the rod-shaped particles was observed in 3.5% of HNSCC and in 0.5% of pOMCs. UVA-1 irradiation of 15 min in combination with 0.2 and 2 µg/ml of ZnO NP dispersion was shown to reduce the vitality of cancer cell lines significantly in comparison to cells without NP exposure or UVA-1 treatment only. For HLaC 78, a significant reduction in viable cells was already seen at 10 min of UVA-1 treatment and a ZnO NP concentration of 2 µg/ml. Flow cytometry indicated that cell death occurred primarily through necrosis. In pOMCs, vitality was not influenced either by UVA-1 treatment or ZnO NP exposure up to 2 µg/ml or a combination of both. ZnO NPs showed cytotoxicity at 20 µg/ml without UVA-1. Due to their photocatalytic properties, ZnO NPs may induce cell death in human HNSCC cell lines in vitro. Further studies will evaluate a possible benefit in adjuvant cancer therapy.

[Tumour regression is not predictive for higher risk of sentinel node involvement in thin melanomas (Breslow thickness < or = 1 mm)]. / La régression tumorale n'est pas un facteur de risque d'atteinte du ganglion sentinelle dans les mélanomes fins (indice de Breslow < or = 1 mm).

BACKGROUND: Thin melanomas (Breslow thickness < or = 1 mm) are considered highly curable. The aim of this study was to evaluate the correlation between histological tumour regression and sentinel lymph node (SLN) involvement in thin melanomas. PATIENTS AND METHODS: This was a retrospective single-centre study of 34 patients with thin melanomas undergoing SLN biopsy between April 1998 and January 2005. RESULTS: The study included 14 women and 20 men of mean age 56.3 years. Melanomas were located on the neck (n=3), soles (n=4), trunk (n=13) and extremities (n=14). Pathological examination showed 25 SSM, four acral lentiginous melanomas, three in situ melanomas, one nodular melanoma and one unclassified melanoma with a mean Breslow thickness of 0.57 mm. Histological tumour regression was observed in 26 over 34 cases and ulceration was found in one case. Clark levels were as follows: I (n=3), II (n=20), III (n=9), IV (n=2). Growth phase was available in 15 cases (seven radial and eight vertical). Mitotic rates, available in 24 cases, were: 0 (n=9), 1 (n=11), 2 (n=2), 3 (n=1), 6 (n=1). One patient with histological tumour regression (2.9% of cases and 3.8% of cases with regressing tumours) had a metastatic SLN. One patient negative for SLN had a lung relapse and died of the disease. Mean follow-up was 26.2 months. CONCLUSION: The results of the present study and the analysis of the literature show that histological regression of the primary tumour does not seem predictive of higher risk of SLN involvement in thin melanomas. This suggests that screening for SLN is not indicated in thin melanomas, even those with histological regression.

Kaposiform hemangioendothelioma arising in the deltoid muscle without the Kasabach-Merritt phenomenon.

Kaposiform hemangioendothelioma (KHE) is a rare tumor that occurs nearly exclusively during infancy and childhood. It has features common to both capillary hemangioma and Kaposi sarcoma and for that reason many terms have been used for these tumors including "Kaposi-like infantile hemangioendothelioma" and "hemangioma with Kaposi sarcoma-like features." KHE typically presents as an ill-defined, red to purple, indurated plaque and is often complicated by the Kasabach-Merritt phenomenon (KMP), a condition of severe thrombocytopenia and consumptive coagulopathy. Knowledge of the radiological findings of this uncommon tumor might be helpful for diagnosis. We present the MRI features of a case of KHE with neither typical skin lesions nor the Kasabach-Merritt phenomenon.

Head-facial hemangiomas studied with scanning electron microscopy.

INTRODUCTION: Hemangiomas of the head or face are a frequent vascular pathology, consisting in an embryonic dysplasia that involves the cranial-facial vascular network. Hemangiomas show clinical, morphological, developmental, and structural changes during their course. METHODS: Morphological, structural, ultrastructural, and clinical characteristics of head-facial hemangiomas were studied in 28 patients admitted in our hospital. Nineteen of these patients underwent surgery for the removal of the hemangiomas, whereas 9 patients were not operated on. All the removed tissues were transferred in our laboratories for the morphological staining. Light microscopy, transmission electron microscopy, and scanning electron microscopy techniques were used for the observation of all microanatomical details. All patients were studied for a clinical diagnosis, and many were subjected to surgical therapy. RESULTS: The morphological results revealed numerous microanatomical characteristics of the hemangiomatous vessels. The observation by light microscopy shows the afferent and the efferent vessels for every microhemangioma. All the layers of the arterial wall are uneven. The lumen of the arteriole is entirely used by a blood clot. The observation by transmission electron microscopy shows that it was impossible to see the limits of the different layers (endothelium, medial layer, and adventitia) in the whole wall of the vessels. Moreover, both the muscular and elastic components are disarranged and replaced with connective tissue. The observation by scanning electron microscopy shows that the corrosion cast of the hemangioma offers 3 periods of filling: initially with partial filling of the arteriolar and of the whole cast, intermediate with the entire filling of the whole cast (including arteriole and venule), and a last period with a partial emptying of the arteriolar and whole cast while the venule remains totally injected with resin. CONCLUSION: Our morphological results can be useful to clinicians for a precise diagnosis on the aftereffects of hemangiomas.

Quantitative analysis of invasive front in tongue cancer using ultrasonography.

PURPOSE: Ultrasonography is a useful diagnostic modality for the head and neck region. The mode of invasion is one of the predictive factors for the cervical lymph node metastasis. The purpose of this study is to evaluate the correlation between quantified invasive fronts from ultrasonographic images and the pathological malignancy grading of tongue squamous cell carcinoma. MATERIALS AND METHODS: Forty-eight previously untreated T1-2N0 tongue squamous cell carcinoma patients were enrolled. The ultrasonographic images of intraoral lesions and the excised lesions were collected. The invasive front on the ultrasonographic image was analyzed by a multipurpose software package. The length of the invasive front and the line of smoothed invasive front were measured and the invasive front ratio was calculated. The hematoxylin and eosin stained sections were assessed according to modified Jakobsson's malignancy grading as low and high malignancy grade groups. The correlation between invasive front ratio and the histological malignancy grade groups was evaluated. RESULTS: The mean invasive front ratio for the excised lesions was 1.145 in low malignancy grade group and 1.248 in high malignancy grade group. These values as determined for the intraoral lesions were 1.074 in the low malignancy grade group and 1.174 in high malignancy grade group. Significant statistical differences were observed between the 2 groups by Mann-Whitney's U test in both images. CONCLUSIONS: Invasive front ratio is related to the histological malignancy grade group. It can be considered to be a valuable diagnostic method which can predict neck node metastasis, and help surgeons to choose adequate neck treatment.

Ultrastructural characteristics of chemodectomas.

We present results of morphological study of chemodectoma samples obtained during surgery. Specific ultrastructural features and protein-synthesizing and proliferative activity of light and dark cells were demonstrated using data of electron radioautography.

Primary cutaneous adenoid cystic carcinoma.

Primary cutaneous adenoid cystic carcinoma is a rare, slow-growing malignancy first described by Boggio in 1975. This tumor characteristically consists of basophilic cells with a distinct adenoid or cribriform pattern in the mid to deep reticular dermis. Modified myoepithelial cells with prominent basement membrane material often surround true lumina. Definitive diagnosis relies on the characteristic histologic features and the exclusion of metastatic disease. We describe two patients who presented with painful papules of the scalp and were successfully treated with wide local excision.

Papillary haemangioma. A distinctive cutaneous haemangioma of the head and neck area containing eosinophilic hyaline globules.

AIMS: To investigate and define a morphologically distinctive group of cutaneous papillary haemangiomas. METHODS AND RESULTS: Eleven patients (seven male, four female, age range 1-77 years, median 57) were identified with a solitary bluish cutaneous papule (median size 11 mm) arising in the head and neck region. Most lesions had been present for several years. None of the patients had associated systemic disease or polyneuropathy, organomegaly, endocrinopathy, M-protein, skin changes (POEMS) syndrome. Only one lesion recurred locally. The lesions showed predominantly intravascular papillary growth within multiple ectatic dermal vessels. The papillae had cellular cores containing pericytes and stromal cells, arranged around normal small capillaries. The surfaces of the papillae were covered by focally swollen endothelial cells containing numerous hyaline globules, ultrastructurally representing giant lysosomes containing organelle debris and fat vacuoles (so-called thanatosomes). These endothelial cells were immunopositive for CD31 and CD34 but negative for D2-40 (podoplanin). CONCLUSIONS: Papillary haemangioma is a distinctive benign cutaneous lesion containing eosinophilic hyaline globules consistent with dysfunction of the autophagocytic-lysosomal pathway.

Cutaneous Langerhans cell sarcoma: a case report and review of the literature.

Langerhans cell sarcoma (LCS) is a neoplastic proliferation of Langerhans cells that occurs in lymph nodes, liver, skin, spleen, lung, and bone. We report a case of LCS in a 47-year-old man with a 6-month history of scalp mass and cervical lymphadenopathy. Clinical and pathologic data were available. A histologic examination demonstrated a proliferation of cells with malignant cytologic features. Because of its poorly differentiated morphologic features, hematologic and nonhematologic entities were ruled out by immunohistochemical screening with a broad panel of antibodies. Ultrastructural studies demonstrating Birbeck granules and consistent expression of CD1a, S-100 protein, and langerin by immunohistochemistry were helpful in identifying the Langerhans cell origin.