Density distribution, cytomorphologic features and immunologic characteristics of ovarian and endometrial clear cell carcinomas

Three subpopulations of cancer cells with different morphologic features were separated by density gradient centrifugation of two ascitic fluids and one cystic fluid from one patient with ovarian clear cell carcinoma and one with endometrial clear cell carcinoma. Immunophenotypic analyses of isolated fractions using polyclonal and monoclonal antibodies against ovarian carcinoma-associated antigens revealed significant immunologic heterogeneity among the tumor cells. The identical histopathologic structures of the ovarian and endometrial clear cell carcinomas and the similar distribution and immunologic reactivity of the cell types isolated from the ascitic and cystic fluids confirmed the common histogenesis of both cancers. These findings suggest that the conventional cytologic diagnosis of clear cell carcinomas could be supplemented by immunofluorescent staining. Density gradient centrifugation appeared to be a useful method for the separation of mesothelial cells.     

Cytokeratins in normal lung and lung carcinomas. I. Adenocarcinomas, squamous cell carcinomas and cultured cell lines

The various epithelial cells of the lower respiratory tract and the carcinomas derived from them differ markedly in their differentiation characteristics. Using immunofluorescence microscopy and two-dimensional gel electrophoresis of cytoskeletal proteins from microdissected tissues we have considered whether cytokeratin polypeptides can serve as markers of cell differentiation in epithelia from various parts of the human and bovine lower respiratory tract. In addition , we have compared these protein patterns with those found in the two commonest types of human lung carcinoma and in several cultured lung carcinoma cell lines. By immunofluorescence microscopy, broad spectrum antibodies to cytokeratins stain all epithelial cells of the respiratory tract, including basal, ciliated, goblet, and alveolar cells as well as all tumor cells of adenocarcinomas and squamous cell carcinomas. However, in contrast, selective cytokeratin antibodies reveal cell type-related differences. Basal cells of the bronchial epithelium react with antibodies raised against a specific epidermal keratin polypeptide but not with antibodies derived from cytokeratins characteristic of simple epithelia. When examined by two-dimensional gel electrophoresis, the alveolar cells of human lung show cytokeratin polypeptides typical of simple epithelia (nos. 7, 8, 18 and 19) whereas the bronchial epithelium expresses, in addition, basic cytokeratins (no. 5, small amounts of no. 6) as well as the acidic polypeptides nos. 15 and 17. Bovine alveolar cells also differ from cells of the tracheal epithelium by the absence of a basic cytokeratin polypeptide. All adenocarcinomas of the lung reveal a "simple-epithelium-type" cytokeratin pattern (nos. 7, 8, 18 and 19). In contrast, squamous cell carcinomas of the lung contain an unusual complexity of cytokeratins. We have consistently found polypeptides nos. 5, 6, 8, 13, 17, 18 and 19 and, in some cases, variable amounts of cytokeratins nos. 4, 14 and 15. Several established cell lines derived from human lung carcinomas (SK-LU-1, Calu -1, SK-MES-1 and A-549) show a uniform pattern of cytokeratin polypeptides (nos. 7, 8, 18 and 19), similar to that found in adenocarcinomas. In addition, vimentin filaments are produced in all the cell lines examined, except for SK-LU-1.(ABSTRACT TRUNCATED AT 400 WORDS)     

Nuclear DNA content, cytomorphologic features and clinical characteristics of small cell carcinoma of the lung

The relationship between the cytomorphologic features, the nuclear DNA patterns and the clinical prognosis of patients with small cell carcinoma of the lung was studied. In cases in the long-survival group (greater than or equal to 24 months), bronchial brushing smears contained a relatively high frequency of nuclei with large, irregular shapes and finely granular chromatin patterns, in comparison with patients in the short-survival group (less than or equal to 9 months); the correlation was not statistically significant, however. The incidence of cells with round or oval nuclei and finely granular chromatin patterns was higher in patients whose cells had hyperdiploid DNA patterns than for patients whose cells had near-diploid patterns; again, the difference was not statistically significant. Patients whose tumor cells had hyperdiploid DNA patterns had significantly shorter survival times than did patients whose tumor cells had near-diploid patterns. These results indicate that (1) judging the nuclear DNA pattern from the cytomorphologic features of small cell carcinoma is unreliable and (2) the nuclear DNA patterns are related to the clinical prognosis of patients with small cell carcinoma of the lung.     

Cytosolic levels of neuron-specific enolase in squamous cell carcinomas of the lung

To study the behavior and possible correlations of neuron-specific enolase (NSE) with other clinicobiological parameters, we measured the cytosolic levels of this marker by means of an immunoradiometric assay (IRMA) in 95 squamous cell lung carcinoma samples. We also analyzed the levels of pS2, tissue-type plasminogen activator (t-PA), hyaluronic acid (HA), free beta subunit of human chorionic gonadotropin (beta-HCG), CYFRA 21.1 and CA 125 in cytosol. On the cell surface we analyzed the concentrations of epidermal growth factor receptor (EGFR), HA, erbB-2 oncoprotein, CD44s, CD44v5 and CD44v6. Other parameters considered were clinical stage, lymph node involvement, histological grade (HG), ploidy and the cellular S-phase fraction measured by flow cytometry on nuclei obtained from fresh tissues. In the 95 squamous cell carcinomas the cytosolic levels of NSE varied from 4.5 to 2235 ng/mg protein (median: 267) and were significantly higher (p < 0.001) than those observed in 38 samples of normal pulmonary tissue obtained from the same patients (range: 56-657; median: 141.5). When classifying tumors according to the different parameters analyzed, we observed that the levels of NSE were higher in aneuploid than in diploid cases (p = 0.046) and in those that were HG3 than in those that were HG2 (p < 0.001). Tumors with high NSE levels (> 422 ng/mg protein; 75th percentile) were more likely to have high S-phase values (p = 0.012) and were more frequently aneuploid (p = 0.038) and HG3 (p < 0.001) than those with low levels of NSE (< 180 ng/mg protein; 25th percentile). These results lead us to the following conclusions: 1) the cytosolic concentrations of NSE are significantly higher in squamous cell carcinomas than in healthy pulmonary tissue, and 2) the cytosolic concentrations of NSE are not correlated with clinical stage or nodal involvement. However, in our study higher levels of the enzyme were statistically correlated with aneuploidy, histological grade 3 and S-phase. This may explain its association with poorer outcome and progression, but also the more favorable response of tumors with elevated NSE to chemotherapy, as suggested by other groups.     

Electronmicroscopic observations on epidermoid (squamous cell) carcinomas of the lung

Five epidermoid (squamous cell) carcinomas of the lung histopathologically diagnosed were ultrastructurally analysed; special attention was given to the poorly differentiated-immature areas of proliferation. Wide intercellular spaces between irregular cytoplasms with protrusions and microvilli, a high incidence of indentations of nuclear membranes, a large amount of nuclear bodies, shape, size and structure anomalies of mitochondria, a great number of desmosomes and of tonofilaments and tonofibril bundles and their relations with desmosomes and with the finger-like cytoplasmic expansions were noticed. A few secretory granules were also present in these poorly differentiated-immature areas of epidermoid carcinomas of the lung.     

Differences in methylation profiles between HPV-positive and HPV-negative oropharynx squamous cell carcinoma

Oropharyngeal squamous cell carcinoma (OPSCC) is associated with human papillomavirus (HPV). HPV-positive OPSCC is considered a distinct molecular entity with a better prognosis than HPV-negative cases of OPSCC. However, the exact pathogenic mechanisms underlying the differences in clinical and molecular behavior between HPV-positive and HPV-negative OPSCC remain poorly understood. Epigenetic events play an important role in the development of cancer. Hypermethylation of DNA in promoter regions and global hypomethylation are 2 epigenetic changes that have been frequently observed in human cancers. It is suggested that heterogeneous epigenetic changes play a role in the clinical and biological differences between HPV-positive and HPV-negative tumors. Unraveling the differences in methylation profiles of HPV-associated OPSCC may provide for promising clinical applications and may pave the road for personalized cancer treatment. This systematic review aims to assess the current state of knowledge regarding differences in promoter hypermethylation and global methylation between HPV-positive and HPV-negative OPSCC.     

Cellular and clinicopathological features of the IL-33/ST2 axis in human esophageal squamous cell carcinomas

Background

Emerging evidence has suggested that interleukin (IL)-33 and its primary functional receptor ST2 are involved in the pathogenesis of tumorigenesis.

Methods

Using immunohistochemistry (IHC) and double immunofluorescence staining, we characterized the cellular and clinicopathological features of the IL-33/ST2 axis in different compartments in human esophageal squamous cell carcinoma (ESCC) surgical specimens.

Results

IHC data revealed an increased expression of IL-33-immunoreactivity (IR) and ST2-IR located in both ESCC cells and tumor stromal cells; which were associated with advanced clinicopathological features such as TNM stages and node involvement. However, the Kaplan–Meier analysis showed that densities of neither IL-33 positive nor ST2 positive cells in both the ESCC mass and stroma were associated with the overall survival rate in patients with ESCC. Double immunofluorescence staining for cellular feature analysis demonstrated that these IL-33 positive and ST2 positive cells in ESCCs were with a high proliferation rate, and IL-33-IR was frequently co-expressed with ST2-IR in both ESCC and stromal cells.

Conclusion

Significant altered cellular features of the IL-33/ST2 axis in ESCCs were associated with advanced clinicopathological variables. The data suggest that the IL-33/ST2 axis might be involved in the progression of human ESCCs.

     

首发为食管鳞癌的多原发癌患者临床特征及生存分析

目的:回顾性分析首发癌为食管鳞癌的多原发癌(ESCCFPM)患者的肿瘤临床特点以及生存预后等临床信息,更好的了解食管鳞癌与其他癌症之间的联系,为指导临床诊治提供相应依据.方法:收集美国国立癌症研究所监测、流行病学和结果数据库(SEER) 2004年1月1日至2016年12月31日ESCCFPM患者临床资料,Kaplan...     

Elevated GAPDH expression is associated with the proliferation and invasion of lung and esophageal squamous cell carcinomas

Glyceraldehyde‐3‐phosphate dehydrogenase, is one of the most investigated housekeeping genes and widely used as an internal control in analysis of gene expression levels. The present study was designed to assess whether GAPDH is associated with cancer cell growth and progression and, therefore may not be a good internal control in cancer research. Our results from clinical tissue studies showed that the levels of GAPDH protein were significantly up‐regulated in lung squamous cell carcinoma tissues, compared with the adjacent normal lung tissues, and this was confirmed by western blotting and immunohistochemistry. GAPDH knockdown by siRNA resulted in significant reductions in proliferation, migration, and invasion of lung squamous carcinoma cells in vitro. In a nude mouse cancer xenograft model, GAPDH knockdown significantly inhibited the cell proliferation and migration/invasion in vivo. In summary, GAPDH may not be an appropriate internal control for gene expression studies, especially in cancer research. The role of GAPDH in cancer development and progression should be further examined in pre‐clinical and clinical studies.     

Correlation between nitric oxide and cyclooxygenase-2 pathways in head and neck squamous cell carcinomas

We investigated the interactions between inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) pathways in head and neck squamous cell carcinomas (HNSCCs) and in two carcinoma cell lines. HNSCCs showed an up-regulation of both pathways which were strongly correlated with each other (p=0.02) and with tumor vascularization (p=0.0001 and p=0.008, respectively). In carcinoma cells, Escherichia coli lipopolysaccharide (LPS) and EGF treatment up-regulated both pathways. NOS inhibitor N(G)-monomethyl-L-arginine methyl ester (L-NAME) inhibited this up-regulation. LPS or EGF induced iNOS expression that was not altered by NOS or COX-2 inhibitors. Conversely, LPS or EGF promoted COX-2 expression that was decreased by L-NAME. The NO donor S-nitroso-acetyl-penicillamine (SNAP) up-regulated COX-2 pathway and this effect was reduced by the guanylate cyclase inhibitor methylene blue. Thus, in squamous carcinoma cells, NO increases the activity of COX-2 pathway and this effect is probably mediated by endocellular cGMP level, with potential implications on tumor growth, angiogenesis, and therapy.     

Correlation between the expression of cyclin A protein and p53 activity in oral squamous cell carcinomas

Cyclins and wild-type p53 protein are prime cell cycle regulators and may be involved in tumorigenesis. Cyclin A is a late S cyclin and its abnormalities have been reported in several cancers, including oral squamous cell carcinomas. To explore whether aberrant G1/S in p53 mutant tumours leads to increased cyclin A protein in oral squamous cell carcinomas (OSCC), a total of 39 samples were evaluated for the expression of cyclin A and p53 protein by an immunohistochemical method using a labelled polymer assay. These samples comprised two hyperkeratotic and three oral premalignant lesions (two moderate and one severe dysplastic lesions), and 27 OSCC, together with seven healthy controls. The results demonstrated that the cyclin A protein was localized and highly expressed in the nuclei of the tumour cells. Although there was no correlation between cyclin A detection and the local lymph node involvement, a positive correlation was noted between the positivity of cyclin A and p53 protein (p <0.05). The results suggested that cyclin A may contribute to the progression of oral cancer and correlated to some degree with that of the p53 gene activity.     

Differences between cell lines of uterine cervical glassy cell carcinoma and large cell nonkeratinizing squamous cell carcinoma.

Cell lines were established from two uterine cervical cancers, a glassy cell carcinoma (GCC) and a large cell nonkeratinizing squamous cell carcinoma (LCSC), and studied by a variety of techniques, including histology, chromosome analysis, heterotransplantation and tumor marker analyses. There were radical differences in the morphology, heterotransplantability, production of tumor markers, etc., between the cultures of these morphologically similar cancers. The LCSC line (HKMUS) consisted of polygonal and round cells containing tonofilaments; these cells discharged tumor antigen-4 (TA-4) into the conditioned media. HKMUS was heterotransplantable into the subcutis of nude mice to form LCSC. On the other hand, the GCC line (HOKUG) consisted of round or spindle-shaped cells. HOKUG was easily transplanted into the subcutis or intraabdominal cavity of nude mice and metastasized easily. It discharged TA-4, carbohydrate antigen 125 (CA125) and neuron-specific enolase (NSE) into the conditioned media. The histologic picture of GCC revealed numerous blood vessels and a rapid proliferation of the cells. GCC, which is considered to be a mixed carcinoma having the characteristics of both squamous carcinoma and adenocarcinoma, seems to be a cancer of unpredictable prognosis as compared to LCSC, possibly due to its rapid proliferation and easy metastasis, leading to peritonitis carcinomatosa.     

Nuclear morphometry in canine acanthomatous ameloblastomas and squamous cell carcinomas

The aim of this study was to evaluate whether morphometrical analysis can be of diagnostic value for canine acanthomatous ameloblastoma. We calculated, by means of an automated image analyser, some morphometric nuclear parameters, in particular: mean nuclear area (MNA), mean nuclear perimeter (MNP), maximum and minimum diameters (MDx and MDm) coefficient of variation of the nuclear area (NACV), largest to smallest dimension ratio (LS ratio), and form factor (FF), in 8 canine acanthomatous ameloblastomas, and we compared these morphometric data to those of 13 squamous cell carcinomas of canine gingiva. The results indicated a progressive increase of the MNA, NACV, MNP and MDm proceeding from acanthomatous ameloblastomas (MNA: 42.11+/-8.74; NACV: 28,36+/-7,23; MNP: 24.18+/- 2.68; MDm: 5.69+/-0.49) to squamous cell carcinomas (MNA:49,69+/-9,10; NACV: 30,89+/-7,75; MNP: 25.63+/-2.54; MDm: 6.64+/-0.73). On the contrary, the LS ratio and the FF resulted greater in acanthomatous ameloblastomas (LS ratio: 1,63+/-0,12; FF: 1,13+/-0,002) than in SCCs (LS ratio: 1,40+/-0,12; FF:0.91+/-0.38). Moreover, the MNA, MNP,MDx and MDm resulted similar (MNA: p=0.89; MNP: p=0,65; MDm: p=0,16; MDx: p=0,13) in a subset of four acanthomatous ameloblastomas with cellular atypia (MNA:49,01+/-6,88; MNP: 26,28+/-1,99; MDm: 6.08+/-0.41; MDx: 10.18+/-0.88) and in squamous cell carcinomas (MNA:49.69+/-9,10; MNP: 25.63+/-2.54; MDm: 6.64+/-0.73; MDx: 9.26+/-1.05). While the NACV values resulted higher in typical acanthomatous ameloblastoma (29,99+/-6,06) than in atypical acanthomatous ameloblastoma (26,74+/-8,84) and similar to those of the SCCs (30,89+/-7,75). These results seem to confirm that acanthomatous ameloblastoma is a malignant or potentially malignant lesion and emphasizes that nuclear morphometry analysis can be an useful diagnostic and prognostic method in canine oral pathology.     

Proliferating activity in oral dysplastic leasions and squamous cell carcinomas

The aim of the study was the quantitative analysis of AgNORs in oral squamous cell carcinomas as well as in dysplastic epithelial changes accompanied and not accompanied by oral squamous cell carcinomas. AgNOR proteins were visualized in histological slides using silver impregnation technique according to D. Ploton. In each sample 100 cell nuclei were assessed. The study used 54 cases of proliferating oral epithelial changes divided into 3 groups: group I consisting of 13 cases of dysplastic lesions not accompanied by oral squamous cell carcinomas; group II (a total of 18 cases) containing dysplastic lesions situated in the vicinity of oral carcinomas and group III (23 cases) with oral squamous cell carcinomas. Statistically significant differences were found between groups with mild dysplasia and groups with severe dysplasia as well as squamous cell carcinomas. Statistical analysis did not show any differences in the number of AgNORs between squamous cell carcinomas and epithelial lesions with severe dysplasia. Our results demonstrate that the analysis of AgNORs expression can serve only as an additional parameter to evaluate the potential of malignant transformation.     

Co-expression of the squamous cell carcinoma antigens 1 and 2 in normal adult human tissues and squamous cell carcinomas.

Squamous cell carcinoma antigen (SCCA) serves as a serological marker for advanced squamous cell carcinomas (SCCs) and as an indicator of therapeutic response. Recent molecular studies show that the SCCA is transcribed by two almost identical tandemly arrayed genes, SCCA1 and SCCA2. These genes are members of the high molecular weight serine proteinase inhibitor (serpin) superfamily. Although SCCA1 and SCCA2 are 92% identical at the amino acid level, they have distinct biochemical properties. Paradoxically, SCCA1 is an inhibitor of papain-like cysteine proteinases, such as cathepsins L, S, and K, whereas SCCA2 inhibits chymotrypsin-like serine proteinases, cathepsin G, and mast cell chymase. Using a new set of discriminatory monoclonal antibodies (MAbs) and polymerase chain reaction (PCR) assay, we showed that SCCA1 and SCCA2 were co-expressed in the suprabasal layers of the stratified squamous epithelium of the tongue, tonsil, esophagus, uterine cervix and vagina, Hassall's corpuscles of the thymus, and some areas of the skin. SCCA1 and SCCA2 also were detected in the pseudo-stratified columnar epithelium of the conducting airways. Examination of squamous cell carcinomas of the lung and head and neck showed that SCCA1 and SCCA2 were co-expressed in moderately and well-differentiated tumors. Moreover, there was no differential expression between these SCCA "isoforms" in normal or malignant tissues. In contrast to previous studies, these data indicated that the expression of SCCA1 and SCCA2 was not restricted to the squamous epithelium and that these serpins may coordinately regulate cysteine and serine proteinase activity in both normal and transformed tissues.