Morphometrical differences between pseudo-epitheliomatous hyperplasia in granular cell tumours and squamous cell carcinomas

AIM: Granular cell tumour (GCT) is a benign tumour which occasionally induces pseudo-epitheliomatous hyperplasia (PEH) of the covering epithelium, mimicking squamous cell carcinoma (SCC) invasion. PEH cells do not show marked atypia, but the presence of pseudoinvasive patterns remains a diagnostic problem since several misdiagnosed cases have been reported. This study investigated objective morphometric criteria to distinguish GCT-PEH from SCC. METHODS AND RESULTS: The complexity of the epithelial connective tissue interface (ECTI) in 57 profiles from nine SCC and 12 GCT-PEH cases was analysed using fractal geometry. Epithelial and non-epithelial cells were segmented using a space partition procedure and analysed morphometrically. GCT ECTI profiles were significantly more complex than those of SCC. The complexity of ECTI quantified by global and local fractal dimensions allowed up to 86% correct discrimination between GCT-PEH and SCC. The cell-wise discrimination between the two entities using cellular morphology was 76% but when the two approaches were combined, the correct discrimination was 96%. CONCLUSIONS: The architectural features of GCT-PEH and SCC show differences which, when quantified, could be used to differentiate the two diagnostic classes. Characterization of these differences may help to elucidate some of the mechanisms of tumour infiltration and metastasis.     

Are epidermal growth factor and transforming growth factor responsible for pseudoepitheliomatous hyperplasia associated with granular cell tumors?

Granular cell tumors (GCT) are uncommon benign neoplasms that have a predilection for the head and neck region. These tumors can frequently be associated with pseudoepitheliomatous hyperplasia (PEH), which in turn may be mistaken for squamous cell carcinoma. Although epidermal growth factors are overexpressed in squamous cell carcinomas of the head and neck, their presence in PEH, especially its relation to GCT, is unknown. We hypothesize that the expression of epidermal growth factor receptor (EGFR), epidermal growth factor (EGF), and transforming growth factor alpha (TGFalpha) in GCT have a role in the development of PEH overlying some GCT. Sections from 13 cases of GCT (five with overlying PEH) were examined histologically and evaluated immunohistochemically using monoclonal antibodies for EGFR, EGF, and TGFalpha. These were compared with nine cases of PEH independent of GCT. Two of five GCT with overlying PEH and two of six GCT without overlying PEH stained positively for TGFalpha. None of the GCT stained with EGFR or EGF. All cases of PEH, whether or not associated with GCT, were reactive for EGFR and EGF. Four of the five cases of PEH overlying GCT stained with TGFalpha. The staining pattern and intensity of all three antibodies were comparable to that of the adjacent normal squamous mucosa. Among the three antibodies, only TGFalpha in GCT appears to be related to the development of PEH. Epidermal growth factor receptor and EGF do not seem to be directly involved. The reason of PEH formation associated with GCT in the absence of growth factors is unknown.     

Pulmonary epithelioid hemangioendothelioma: a tumor presented as a single cavitary mass

Pulmonary epithelioid hemangioendothelioma (PEH) is a rare tumor that occurs among young women and typically presents as bilateral multiple nodules. In the present report, we describe an uncommon case of PEH presented as a single cavitary nodule in a 33-yr-old asymptomatic man. This is the first case of PEH presented as a single cavitary nodule in the English literature. Three years of the follow-up without treatment was performed. Overall histologic findings were accord with conventional PEH, but some atypical features such as, increased mitotic activity (mean; two per ten high power fields), necrosis, spindling, and pleural and vascular invasion were recognized. Immunohistochemically, the tumor cells were positive for CD34. This report may contribute to the data on clinical findings and natural history of this rare tumor.     

Malignant pulmonary epithelioid hemangioendothelioma with hilar lymph node metastasis

Few cases each of malignant pulmonary epithelioid hemangioendothelioma (PEH) and PEH with lymph node metastasis have been reported. Here we report a case of PEH with lymph node metastasis. A Japanese woman was found to have a 2-cm-diameter mass with small satellite nodules in the right upper lobe of the lung. Microscopic examination revealed solid destructive growth of the main tumor, with epithelioid cells showing cytologic atypia and 3 mitotic figures per 10 high-power fields. Some of the tumor cells had intracytoplasmic lumina that appeared as vacuoles. These lumina were negative for alcian blue and periodic acid Schiff, and contained erythrocytes. However, erythrocytes were seen more frequently within small but distinct vascular channels that were arranged diffusely in the periphery of the main tumor. Other satellite nodules showed conventional PEH morphology. In hilar lymph nodes, the tumor cells resembled those of the main tumor. The vascular origin of the main tumor and satellite nodules was demonstrated by positive immunoreactivity for some endothelial markers. Although the diagnostic features of malignant PEH are not clear, those for PEH in other organs have included nuclear atypia, many mitoses, presence of necrosis, large tumor size, and spindle cell proliferation. The present case met these criteria, except for large tumor size and spindle cell proliferation. In conclusion, atypical cytologic features, the presence of necrosis, a high Ki-67 labeling index, and accompanying nodules of conventional PEH in the same pulmonary lobe suggest that this case was a malignant PEH with hilar lymph node metastasis.     

Intravascular papillary endothelial hyperplasia of the neck masquerading as malignancy on fine-needle aspiration cytology

Papillary endothelial hyperplasia (PEH) is an exuberant, usually intravascular endothelial proliferation that, in many respects, mimics angiosarcoma. A case of PEH originally suggestive of embryonal carcinoma by fine-needle aspiration is presented. A 12-year-old boy presented with a palpable mass on the right side of the neck. The mass was subsequently aspirated. Cytopathologic features showed cohesive sheets of polygonal pleomorphic cells with vesicular nuclei and prominent multiple nucleoli in a hemorrhagic background. Cytologic findings were strongly suggestive of metastatic embryonal carcinoma. There was no evidence of a primary lesion. After the mass was surgically excised, the pathologic findings showed PEH. A retrospective immunocytochemical stain for factor VIII-related antigen on a destained ethanol-fixed smear confirmed the endothelial nature of the polygonal cells. A vascular lesion should be considered, especially when atypical polygonal cells in a hemorrhagic background are present, as they were in this case.     

Peripheral vs central squamous cell carcinoma of the lung. A comparison of clinical features, histopathology, and survival

We reviewed the clinical features and histopathologic findings of 21 peripheral pulmonary squamous cell carcinomas (SCC) resected at our institution between 1961 and 1981 and compared them with 19 central SCCs. Histologic features were scored semiquantitatively from 0 to 3+. Peripheral SCC represented 16% of all resected SCCs. The proportion of patients with multiple symptoms was lower and survival during the 5 years after surgery was better in the peripheral group. Tumor size, mitoses per high-power field (2.4 +/- 0.3 vs 4.1 +/- 0.6 [SEM]), prevalence of lymphatic invasion (19% vs 58%), and lymph node metastases (5% vs 37%) were lower, while chest wall invasion was more frequent (25% vs 0) for peripheral SCCs. Peripheral tumors also had more intense (2 or 3+) lymphoplasmacytic (86% vs 47%) and desmoplastic (95% vs 68%) reactions. Cox regression analysis did not support a significant relationship between tumor location and survival. We conclude that, compared with the central SCC, peripheral SCC is associated with fewer symptoms at presentation and better survival. Morphologically, peripheral SCCs are smaller, have fewer mitoses, less prevalent lymphatic invasion, and a more intense stromal reaction. Improved survival in patients with peripheral SCC may be due to a more favorable stage at the time of initial treatment.     

Slug is associated with poor survival in squamous cell carcinoma of the lung

We investigated the expression of slug in a large set of lung squamous and adenocarcinomas to determine common or dissimilar features in its expression in these two most common forms of lung cancer. To investigate slug related tumor spread we studied the expression of vimentin, claudin 1, MMP2 and MMP9 in these tumors and their relation to slug. Addition, cell invasion assays, mRNA analysis and zymographic tests were performed to study epitheliomesenchymal transition (EMT) related changes in slug blocked lung cell lines. According to the results slug expression did not significantly differ between squamous (SCC) and adenocarcinoma (AC) (P = 0.25). In SCC, slug associated with vimentin (P = 0.016). In AC, claudin 1 associated with MMP2 (P = 0.037). In SCC slug expression had a poor prognositic significance (P = 0.006) and it had independent prognostic value (P = 0.037). In AC MMP2 had a worsening impact on survival (P = 0.021) and it had independent prognostic value (P = 0.002). In cell invasion assays, slug knockdown inhibited the invasion and migration of BEAS-2B, SK-LU1 and SK-MES1 cell lines. The mRNA expression of claudin 1 was downregulated in SK-LU1 cell line. Both tumor cell lines expressed MMP2 and in SK-MES1 slug inhibited line MMP2 appeared to decrease. The results show that slug associated EMT is more pronounced in lung SCC than AC. Slug associated with vimentin in SCC and had an independent prognostic value in this tumor type. Forced slug inhibition might be one putative way of treatment of SCC of the lung.     

Pyogenic granuloma of the oral cavity: comparative study of its clinicopathological and immunohistochemical features

There are two histological types of pyogenic granuloma (PG) of the oral cavity: the lobular capillary hemangioma (LCH) and non-LCH type. The aim of the present study was to examine and compare the clinical features, etiological factors, diameter of vascular elements and immunohistochemical features of LCH and non-LCH histological types of PG to determine whether they are two distinct entities. Thirty cases of LCH and 26 cases of non-LCH PG were retrieved and retrospectively studied. Clinically, LCH PG occurred more frequently (66.4%) as sessile lesion whereas non-LCH PG occurred as pedunculated (77%). Non-LCH PG was associated more frequently (86.4%) with etiological factors. The lobular area of the LCH PG contained a greater number of blood vessels with small luminal diameter than did the central area of non-LCH PG. In the central area of non-LCH PG a significantly greater number of vessels with perivascular mesenchymal cells non-reactive for alpha-smooth muscle actin and muscle-specific actin was present than in the lobular area of LCH PG. The differences found in the present study suggest that the two histological types of PG represent distinct entities.     

Ultrastructural analysis of small cell carcinomas of the ovary

Small cell carcinoma (SCC) of the ovary is a rare tumor of young females, frequently associated with hypercalcemia. Eight SCC specimens from six patients, including four with hypercalcemia, were examined by electron microscopy to determine if certain ultrastructural features were consistently present to be of diagnostic value and to assess histogenesis. Each tumor had epithelial features with frequent desmosome-like junctions and partial investment with basal lamina. The most consistent and prominent finding was abundant dilated rough endoplasmic reticulum (RER) forming large vesicles filled with homogeneous granular (proteinaceous) material of variable density. RER vesicles were identified in all six primary tumors and in both recurrent and metastatic lesions. Dense-core granules of neurosecretory type were absent. None showed immunohistochemical reactivity for parathyroid hormone. The ultrastructural features of SCC are sufficiently constant to be of diagnostic value in the differential diagnosis of ovarian neoplasms. Histogenesis remains obscure.     

Clonal chromosome abnormalities in premalignant lesions of the skin

Two lesions, actinic keratosis (AK) and squamous cell carcinoma in situ (CIS), are believed to be precursors of squamous cell carcinoma (SCC) of the skin. These lesions can serve as an excellent model system for studying genetic changes associated with the inception of skin SCC. In the present study, five such lesions of the skin, three AKs and two AK+CIS, from three patients were short-term cultured and analyzed cytogenetically. One of the patients (case 3) had also an SCC in addition to three premalignant lesions. All lesions, but one, showed clonal karyotypic abnormalities. The recurrent changes identified were numerical, that is, +7 and +20. The structural rearrangements found in three AK were different, but it could be noted that the distal part of the long arm of chromosome 4 was involved in two AK and the SCC of case 3A. It was also interesting that chromosome 1 participated in structural rearrangements in three AK with band 1p31 being involved in two tumors. The karyotypic profile of these lesions is compared with that of skin SCC; it turns out that the general patterns are different in the sense that the SCC more often have complex karyotypes and display unbalanced aberrations involving the centromeric regions. Some karyotypic similarities between the SCC and their precursors are revealed. The fact that the structural rearrangements involving chromosomal band 3p13 and the centromeric region of chromosome 3 in AK are common features for many types of malignant tumors, including skin SCC, indicates that these changes are early genetic events associated with malignant transformation.     

The magnitude and duration of post-exercise hypotension after land and water exercises

The objective of the study was to determine and compare the magnitude and duration of post-exercise hypotension (PEH) during free-living conditions after an acute session of concurrent water and land exercise in individuals with prehypertension and hypertension. Twenty-one men and women (aged 52?±?10?years) volunteered for the study. Participants completed a no exercise control session, a water exercise session and a land exercise session in random order. After all three sessions, participants underwent 24-h monitoring using an Ergoscan ambulatory BP monitoring device. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were monitored to determine changes from resting values after each session and to compare the PEH responses between land and water exercises. During daytime, both land and water exercises resulted in significantly lower SBP (12.7 and 11.3?mmHg) compared to the control session (2.3?mmHg). The PEH response lasted for 24?h after land exercise and 9?h after water exercise. There was no difference in the daytime DBP for the three treatments (P?>?0.05). Although all three groups showed significant reductions during nighttime, both exercise treatments showed greater nocturnal falls in BP than the control treatment. This is the first study to show that the magnitude of the PEH response is similar for land and water exercises, although the duration of PEH may be longer for land exercise. These results suggest that water exercise is a safe alternative exercise modality for individuals with suspected and known hypertension.     

Histological subtypes of oral non-tonsillar squamous cell carcinoma in dogs

Several histological subtypes and grades of oral squamous cell carcinoma (SCC) are described in human literature and these subtypes have distinct morphological features and biological behaviour. This retrospective study (1990-2010) included 84 dogs diagnosed with SCC of the oral cavity and oropharynx, excluding the tonsils. Sixty-nine of the SCCs (82.1%) were further diagnosed as conventional SCC (CSCC) (33 [47.8%] well-differentiated, 31 [44.9%] moderately-differentiated and five [7.3%] poorly-differentiated), five (5.95%) each as papillary SCC and basaloid SCC, three (3.6%) as adenosquamous carcinoma and two (2.4%) as spindle cell carcinoma. Compared with the general hospital population, neutered female dogs, dogs aged 10 to <15 years, English springer spaniels and Shetland sheepdogs were overrepresented. The majority (78.1%) of SCCs were proliferative with or without associated ulceration, although no significant association was observed between the gross appearance and different SCC subtypes. 71.4% of SCCs were located in dentate jaws; however, well-differentiated CSCC more often affected the tongue and other non-dentate mucosal surfaces (P=0.0022). No significant association was found between any of the SCC subtypes and tumour-associated inflammation (TAI), perineural and lymphovascular invasion (PNI, LVI), or between gross appearance of the tumour and tumour location, PNI, LVI or TAI or PNI, LVI, TAI and tumour location.     

Eosinophilia is a favorable prognostic marker for oral cavity and lip squamous cell carcinoma

Eosinophils are frequently encountered with squamous cell carcinomas (SCC) and it has been proposed that tumor‐associated tissue eosinophilia (TATE) could be of prognostic significance in oral SCC. The aim was to evaluate TATE in 83 oral cavity and 16 lip SCCs as well as the best possible use of TATE as a prognostic marker. The number of eosinophils was counted per high power fields (HPF, ×400) in three different representative areas of the tumor and its stroma. The degree of TATE was analyzed in relation to clinicopathological features of tumors and patients’ survival (follow‐up mean 40.7 months) using Fisher's exact test. TATE was detected in 58 (70%) oral and 8 (50%) lip SCC samples. The median number of eosinophils between oral and lip SCC was different (p = 0.028) but TATE was similar per HPF (p = 0.085). Totally, 6% of lip and 21% of oral SCC patients died during the follow‐up. The patients with the higher TATE had significantly better survival than the patients with the lower TATE (p = 0.0136). The best cut‐off value predicting the survival was 4 eosinophils/HPF. TATE is a prognostic marker for oral and lip SCC: more than 4 eosinophils/HPF may predict more favorable prognosis.     

Prognostic significance and effect of chemoradiotherapy on microvessel density (angiogenesis) in esophageal Barrett's esophagus-associated adenocarcinoma and squamous cell carcinoma.

Previous studies have shown that intratumoral microvessel density (IMD) correlates with clinical outcome in a variety of human neoplasms, such as those that arise in the breast, colon, and stomach, suggesting that angiogenesis is important in cancer progression. The aims of this study were to evaluate the prognostic utility of IMD in esophageal Barrett's-associated adenocarcinoma (AdCa) and squamous cell carcinoma (SCC), and to determine the effect of preoperative chemoradiotherapy (chemrad) on this process. Tissue sections of tumor from 67 patients with esophageal carcinoma (45 with Barrett's-associated AdCa, 22 with SCC) were stained with the vascular marker CD31. The IMD was calculated by evaluating at least 5 different 200 x fields of tumor hot spot areas to obtain the mean microvessel count (MVC). The data then were correlated with the clinical and pathological features, chemrad status, and patient survival. The MVC was significantly higher in AdCa (143 +/- 63.2) compared with SCC (77.2 +/- 38.6, P = 0.0001). In AdCa, no correlation was noted between the MVC and any of the clinical or pathological features, including chemrad status. In contrast, in SCC, a statistically significant higher MVC was detected in patients who did not receive chemrad (97.2 +/- 37.3) compared with those who did (48.3 +/- 15.9, P = .002) and in tumors that were larger in size (P = .02). However, the MVC did not correlate with survival in either AdCa or SCC (P > .05). The degree of angiogenesis is not a significant prognostic indicator in either esophageal AdCa or SCC. Preoperative chemrad has a positive effect on reducing the degree of angiogenesis in esophageal carcinoma, particularly SCC.     

Expression of cell cycle-regulatory proteins, MIB-1, p16, p53, and p63, in squamous cell carcinoma of conjunctiva: not associated with human papillomavirus infection

Squamous cell carcinoma (SCC), the most common primary malignant tumor of the conjunctiva, has a variable clinical presentation and immunohistochemical profile. Abundant cell cycles exist, including MIB-1 (Ki67 antigen), p16, p53, and p63, within the conjunctiva SCC. This investigation first reports the expressions of cell cycle markers in SCC. A retrospective study was conducted between December 1976 and June 2004, comprising 13 consecutive patients with conjunctiva SCC who were treated with surgical excision. Detailed clinical parameters were also reviewed. Overexpression of MIB-1, p16, p53, and p63 genes were studied by immunohistochemistry. Genechip containing 39 subtypes was used to elucidate human papillomavirus (HPV). The study group contained 13 (100%) men, with a mean age of 68±18 years and follow-up period of 20±17 months. The sample included four (33%) SCC located in the left eye and two (17%) recurrent SCC. Overexpression of the p53 and p63 was considerably higher than that of the p16 (P<0.01). HPV DNA was not detected in any of the 13 cases. This work first examined the immunohistochemical overexpression of cell cycle (MIB-1, p16, p53, and p63) in SCC. This investigation then showed that the expression of cell cycles in SCC was associated with key tumor clinicopathological features. This approach can help distinguish the potential roles of cell cycle in the development of SCC.     

Phenylethylidenehydrazine, a novel GABA-transaminase inhibitor, reduces epileptiform activity in rat hippocampal slices

Phenylethylidenehydrazine (PEH), an analog of the monoamine oxidase inhibitor, beta-phenylethylhydrazine (phenelzine), inhibits the gamma-aminobutyric acid (GABA) catabolic enzyme GABA-transaminase and increases brain levels of GABA. GABA is the predominant fast inhibitory transmitter counteracting glutamatergic excitation, and increased neural GABA could influence a wide range of synaptic and circuit properties under both physiologic and pathophysiologic conditions. To examine the scope of these effects, we applied PEH (or vehicle) to rat hippocampal slices and measured basal glutamatergic transmission, synaptic plasticity, and epileptiform activity using extracellular field and whole cell patch clamp recordings. In vitro pre-treatment with PEH (100 microM) increased the GABA content of hippocampal slices by approximately 60% over vehicle-treated controls, but it had no effect on basal field excitatory postsynaptic potentials, tonic GABA currents, paired-pulse facilitation, or long-term potentiation. In contrast, pre-incubation with PEH caused a dose- and time-dependent reduction in epileptiform burst frequency induced by superfusion with Mg2+-free or high-K+ artificial cerebrospinal fluid. Thus, the inhibitory effects of PEH are state-dependent: hyper-excitation during epileptiform bursting was reduced, whereas synaptic transmission and plasticity were unaffected.     

High-resolution copy number and gene expression microarray analyses of head and neck squamous cell carcinoma cell lines of tongue and larynx

Gene amplifications and deletions are frequent in head and neck squamous cell carcinomas (SCC) but the association of these alterations with gene expression is mostly unknown. Here, we characterized genome-wide copy number and gene expression changes on microarrays for 18 oral tongue SCC (OTSCC) cell lines. We identified a number of altered regions including nine high-level amplifications such as 6q12-q14 (CD109, MYO6), 9p24 (JAK2, CD274, SLC1A1, RLN1), 11p12-p13 (TRAF6, COMMD9, TRIM44, FJX1, CD44, PDHX, APIP), 11q13 (FADD, PPFIA1, CTTN), and 14q24 (ABCD4, HBLD1, LTBP2, ZNF410, COQ6, ACYP1, JDP2) where 9% to 64% of genes showed overexpression. Across the whole genome, 26% of the amplified genes had associated overexpression in OTSCC. Furthermore, our data implicated that OTSCC cell lines harbored similar genomic alterations as laryngeal SCC cell lines We have previously analyzed, suggesting that despite differences in clinicopathological features there are no marked differences in molecular genetic alterations of these two HNSCC sites. To identify genes whose expression was associated with copy number increase in head and neck SCC, a statistical analysis for oral tongue and laryngeal SCC cell line data were performed. We pinpointed 1,192 genes that had a statistically significant association between copy number and gene expression. These results suggest that genomic alterations with associated gene expression changes play an important role in the malignant behavior of head and neck SCC. The identified genes provide a basis for further functional validation and may lead to the identification of novel candidates for targeted therapies. This article contains Supplementary Material available at http://www.interscience.wiley.com/jpages/1045-2257/suppmat.     

Malignant transformation of mature cystic teratoma to squamous cell carcinoma involves altered expression of p53‐ and p16/Rb‐dependent cell cycle regulator proteins

Ovarian mature cystic teratomas (MCT) uncommonly undergo malignant transformation to squamous cell carcinoma (SCC). While alterations in the p53 tumor suppressor gene and protein have been shown, few studies have analyzed other molecular changes leading to this malignant conversion. The purpose of the present study was to investigate 21 samples of SCC arising in MCT for altered expression in known p53‐ and p16/Rb‐dependent cell cycle regulatory proteins, and the association between their expression and cellular proliferation and histological features. Overexpression of the p53 protein was observed in 14 SCC (67%), while four (19%) had point mutations in the p53 gene. Reduced expression of the p16 protein was observed in 18 SCC (86%), while p16 gene alterations (hypermethylation (29%) and point mutation (33%)) were found in 11 (52%). Furthermore, a statistically significant correlation was observed between p53 and Rb overexpression (P = 0.0010), and the overexpression of both p53 and Rb was respectively significantly correlated with increased cellular proliferation. The results indicate that alterations in both the p53 and p16‐Rb pathways are associated with SCC arising in MCT.     

A case report of papillary thyroid carcinoma dedifferentiated to squamous cell carcinoma presenting as a lung metastasis: A potential diagnostic pitfall

Papillary thyroid carcinoma (PTC) tall cell variant (TCV) with squamous dedifferentiation is a rare entity. We present a case of 90‐year‐old woman who initially had a 2.8 cm conventional PTC in right lobe of thyroid who, couple decades later, had metastatic dedifferentiated PTC to right neck lymph nodes level II and IV with tall cell features; to right level IV and V lymph nodes with tall cell and squamous components, which recently presented exclusively as squamous cell carcinoma (SCC) metastasizing to lung. The squamous component in the lymph node and SCC in the lung were both positive for squamous marker p63 and PTC markers TTF1, PAX‐8 and BRAF V600E while negative for thyroglobulin and p16. The papillary component was positive for TTF‐1, BRAF V600E and P63 (majority); negative for thyroglobulin and p16. Final diagnoses were rendered based on combination of cytological features and immunohistochemical profiles. This report highlights the utilization of current biomarkers to distinguish between metastatic dedifferentiated PTC with squamous features and primary lung SCC, as well as the importance of recognizing this rare entity.