Elevated expression of nuclear protein kinase CK2α as a poor prognosis indicator in lymph node cancerous metastases of human thyroid cancers.

AIM: To investigate the expression of protein kinase CK2α (CK2α) in human thyroid disease and its relationship with thyroid cancer metastasis. MATERIALS AND METHODS: Using immunohistochemistry we measured the expression of CK2α in 76 benign and malignant human thyroid cancer tissues, including 10 pairs of papillary carcinoma tissues with or without lymph node cancerous metastasis and similarly 10 pairs of lymph nodes. RESULTS: The expression of CK2α was found to be higher in thyroid carcinoma cases (papillary carcinoma, follicular carcinoma, anaplastic carcinoma and medullary carcinoma) than in ones such as chronic lymphocytic thyroiditis, nodular goiter and adenoma. These findings were also confirmed by RT-PCR and Western blotting. More strikingly, elevated expression of CK2α in thyroid papillary carcinoma tissues was not only significantly associated with lymph node cancerous metastasis and clinical stage of thyroid cancers; but also correlated with epithelial-mesenchymal transition (EMT) and high tenascin C (TNC) expression. In addition, EMT and high TNC expression in thyroid carcinoma tissues was significantly associated with lymph node cancerous metastasis. CONCLUSIONS: Elevated expression of nuclear CK2α is a poor prognosis indicator in lymph node cancerous metastasis of human thyroid cancers.

Gene expression and clonality analysis of the androgen receptor and phosphoglycerate kinase genes in polygonal cells and cuboidal cells in so-called pulmonary sclerosing hemangioma.

The histogenesis of polygonal cells and cuboidal cells in so-called pulmonary sclerosing hemangioma remains unclear. To understand their histogenesis, polygonal and cuboidal cells were obtained from pulmonary sclerosing hemangioma tissue using a laser capture microdissection technique. Genomic DNA and total RNA were extracted and mRNA levels of cytokeratin, epithelial membrane antigen, vimentin, surfactant protein B, thyroid transcription factor-1, synaptophysin, and chromogranin-A were analyzed by RT-PCR. DNA was digested with the methylation-sensitive enzymes HhaI or HpaII, followed by nested PCR of the androgen receptor and phosphoglycerate kinase genes. Samples with polymorphisms were identified and a clonality analysis was performed. The cytokeratin, epithelial membrane antigen, and surfactant protein B genes were clearly expressed in cuboidal cells, while the vimentin and synaptophysin genes were clearly expressed and the epithelial membrane antigen gene was weakly expressed in polygonal cells. Thyroid transcription factor-1 was expressed in both cell types, while neither cell type expressed chromogranin-A. Clonality analysis showed the same loss of allele in both cell types (clonality ratio=0) or an unbalanced methylation pattern (clonality ratio<0.25). Polygonal and cuboidal cells in pulmonary sclerosing hemangioma exhibited a uniform pattern of monoclonality, indicating that both cell types are highly likely to originate from a common precursor. The differences in their morphological phenotype might result from their different mature status.

Expression of E-cadherin, beta-catenin and p120ctn in the pulmonary sclerosing hemangioma.

BACKGROUND: The major two types of cells in pulmonary sclerosing hemangiomas (PSH) may be not equally maturity, but this viewpoint needs more evidences. AIM: To determine E-cadherin, beta-catenin and p120(ctn) expression phenotype in cuboidal and polygonal cells, which are the two major cell types in pulmonary sclerosing hemangiomas. METHODS: Specimens were obtained from 25 patients with PSH and 8 patients with pulmonary inflammatory pseudotumors. The expression levels of E-cadherin, beta-catenin and p120(ctn) were detected using a streptavidin peroxidase (SP) immunohistochemical method. RESULTS: E-cadherin, beta-catenin and p120(ctn) were expressed strongly on the cuboidal cell membranes, while beta-catenin was also expressed the cuboid cytoplams in 25 PSH patients. However, in the polygonal cell membranes, the expression levels of these molecules were decreased, and mainly cytoplamic. Specifically, E-cadherin, beta-catenin and p120(ctn) were expressed in both the cytoplasm and on the cell membranes in the intracavitary lining cells of the hemorrhagic regions. The expression phenotype in proliferating type II pneumocytes in the eight pulmonary inflammatory pseudotumors was similar to that in the cuboidal cells in PSH patients. CONCLUSION: The cuboidal cells, resembling inflammatory proliferative type II pneumocytes, display several characteristics of epithelial cells, including normal expression of E-cadherin and catenin. Comparatively, polygonal cells are not as mature as cuboidal cells and lack of expression of E-cadherin and catenin.

[Expression of connexin 43 in lung cancer and its correlation with E-cadherin].

OBJECTIVE: To investigate the expression of connexin 43 and E-cadherin in lung cancer and to study the interaction between the two molecules. METHODS: The expression and correlation of connexin 43 and E-cadherin were evaluated by immunohistochemistry (S-P method) in 85 samples of primary squamous cell carcinoma and adenocarcinoma of the lung. In addition, connexin 43 expression vector was transfected into the lung giant cell carcinoma cell line LH(7) followed by analyses of connexin 43 and E-cadherin expressions, the growth rates and cell cycle profiles of the transfected cells. RESULTS: Comparing with the adjacent non-neoplastic lung tissue, expression of connexin 43 and E-cadherin was decreased in a correlative fashion in both squamous cell carcinomas and adenocarcinomas. Their expression reversely correlated to the degree of tumor cell differentiation, P-TNM stage, and status of lymph note metastasis. The expression of connexin 43 and E-cadherin increased significantly after transfection of connexin 43 expression vector into the LH(7) cells (P < 0.05). Both expressions were limited in the cytoplasm before or after the transfection. The proliferation rate of LH(7) cells was significantly decreased by connexin43 expression (P < 0.05), along with an increase of cell population at G(1) phase and a decrease of percentage of cells in S and G(2) phases (P < 0.05). CONCLUSIONS: Squamous cell carcinoma and adenocarcinoma of lung have a low level of connexin 43 and E-cadherin expression, which are correlated with the clinicopathologic features of the tumors. Transfection expression of connexin 43 gene induces an E-cadherin overexpression and an inhibition of LH(7) cell proliferation indicating the significant role of onnexin 43 in the regulation of cell proliferation.

Expression of serine threonine kinase 15 is associated with poor differentiation in lung squamous cell carcinoma and adenocarcinoma.

Serine threonine kinase 15 (STK15, also named BTAK, Aurora-A, aurora-2, or AIKI) is a type of mitotic kinase. The overexpression of STK15 is significantly associated with carcinogenesis in many tumors. The purpose of the present study was to investigate the expression of STK15 in lung squamous cell carcinoma and adenocarcinoma and analyze the correlation between STK15 expression and clinicopathological factors. The expression patterns of STK15 were examined by immunohistochemistry in 80 lung squamous cell carcinomas and adenocarcinomas and 20 normal lung tissues. The protein and mRNA expression of STK15 were evaluated by western blot and reverse transcription-polymerase chain reaction (RT-PCR) in 40 lung cancer samples and corresponding normal lung tissues. Immunohistochemically, the positivity of STK15 expression was 68.75% (55/80). The STK15 expression was significantly higher in poorly differentiated lung cancers than in well-differentiated or moderately differentiated lung cancers (P = 0.011). Western blot and RT-PCR showed that the protein and mRNA expression of STK15 were correlated (P = 0.044) and significantly higher in tumors than in corresponding normal lung tissues (P < 0.05). These results indicate that the overexpression of STK15 contributes to the carcinogenesis and de-differentiation of lung cancers.

[Study of androgen receptor and phosphoglycerate kinase gene polymorphism in major cellular components of the so-called pulmonary sclerosing hemangioma].

OBJECTIVE: To study the clonality of polygonal cells and surface cuboidal cells in the so-called pulmonary sclerosing hemangioma (PSH). METHODS: 17 female surgically resected PSH were found. The polygonal cells and surface cuboidal cells of the 17 PSH cases were microdissected from routine hematoxylin and eosin-stained sections. Genomic DNA was extracted, pretreated through incubation with methylation-sensitive restrictive endonuclease HhaI or HpaII, and amplified by nested polymerase chain reaction for X chromosome-linked androgen receptor (AR) and phosphoglycerate kinase (PGK) genes. The length polymorphism of AR gene was demonstrated by denaturing polyacrylamide gel electrophoresis and silver staining. The PGK gene products were treated with Bst XI and resolved on agarose gel. RESULTS: Amongst the 17 female cases of PSH, 15 samples were successfully amplified for AR and PGK genes. The rates of polymorphism were 53% (8/15) and 27% (4/15) for AR and PGK genes respectively. Polygonal cells and surface cuboidal cells of 10 cases which were suitable for clonality study, showed the same loss of alleles (clonality ratio = 0) or unbalanced methylation pattern (clonality ratio < 0.25). CONCLUSIONS: The polygonal cells and surface cuboidal cells in PSH demonstrate patterns of monoclonal proliferation, indicating that both represent true neoplastic cells.

[Significance of caveolin-1 expression in primary lung cancer].

OBJECTIVE: To study the expression of caveolin-1 in primary lung cancer and its relationship with microvessel density and clinicopathologic parameters. METHODS: Immunohistochemical study for caveolin-1 and CD34 was performed on paraffin sections of 154 cases of primary lung cancer and adjacent non-neoplastic lung parenchymal tissue, as well as 36 cases with nodal metastasis. Microvessel density was analyzed by CD34 immunostaining. Western blot assay was also employed in tumor and non-neoplastic lung tissues of the 50 cases (25 cases of pulmonary squamous cell carcinoma and 25 cases of pulmonary adenocarcinoma) with fresh specimens available. RESULTS: Immunohistochemical study showed that non-neoplastic bronchial and alveolar epithelium was positive for caveolin-1 (membranous and cytoplasmic). The expression rate of caveolin-1 in lung cancer was 59.1%, which was significantly lower than that in normal lung tissues (P < 0.01). Western blot assay confirmed that the expression of caveolin-1 in pulmonary squamous cell carcinoma and adenocarcinoma was lower than in surrounding non-neoplastic lung tissues (P < 0.01). Caveolin-1 expression in pulmonary small cell carcinoma (7.1%) was significantly lower than that in non-small cell carcinoma (64.3%) (P < 0.01). Within the group of non-small cell carcinoma, the expression of caveolin-1 was much higher in patients with lymph node metastasis (P = 0.005). The expression was also higher in stage III and IV than in stage I and II disease (P = 0.042). CONCLUSIONS: The expression of caveolin-1 is lower in lung cancer tissues than that in non-small cell carcinoma, it is also significantly correlated with tumor stage and lymph node metastasis. Caveolin-1 may play some role in the progression of pulmonary non-small cell carcinoma.