Invasive meningioma is associated with a low expression of E-cadherin and beta-catenin.

Invasive meningioma shows benign histological features (WHO grade 1) and the brain expansion at the tumor-brain interface, and recurs more frequently than common meningiomas. To determine the mechanism of brain expansion, we studied the relationship between invasive meningioma and cell adhesion molecules. Immunostaining for E-cadherin (E-CH), N-cadherin (N-CH), beta-catenin, and Ki-67 was performed in 103 meningiomas that consisted of 61 meningothelial meningiomas, 25 fibrous meningiomas, 12 invasive meningiomas and 5 anaplastic meningiomas. All tumors were negative for N-CH. All the 61 meningothelial meningiomas, 10 of 12 invasive meningiomas, and 3 of 5 anaplastic meningiomas were positive for both E-CH and beta-catenin, while these were both negative in all of the fibrous meningiomas. In invasive meningiomas, the expansive part of the tumor showed a lower rate (4/12 tumors) of E-CH and beta-catenin positivity, while the central part showed a higher rate (10/12 tumors). The Ki-67 labeling index was higher in invasive and anaplastic meningiomas than in meningothelial meningiomas. These results suggest that a reduction in cell adhesion molecules and increased proliferative activity may be related, which may lead to a better understanding of the mechanism of meningioma expansion in the future.

Co-expression of N-cadherin and alpha-fetoprotein in stomach cancer.

Although N-cadherin is necessary for organ formation originating in the endoderm, the expression of N-cadherin in gastric carcinoma and its role has not yet been reported. The present study was conducted to determine the pattern of immunohistochemical expression of E-cadherin and N-cadherin, using formalin-fixed, paraffin-embedded tissues from 97 primary gastric carcinomas, including 17 which were producing alpha-fetoprotein (AFP). Samples were subdivided into 50 tubular adenocarcinomas and 47 poorly differentiated adenocarcinomas. Results showed that E-cadherin was expressed in varying degrees in areas of cell adhesion between tumor cells, in 94 out of 97 cases studied. Three cases which showed no expression of E-cadherin were diagnosed as AFP-producing tumors by immunohistochemistry. Expression of N-cadherin was observed in varying degrees in the intercellular spaces between tumor cells in 11 tubular adenocarcinomas and in six poorly differentiated adenocarcinomas, including E-cadherin-negative cases, all of which were AFP positive. The present findings suggest a possible role for N-cadherin in gastric carcinoma.

Comparison of the DNA extraction methods for polymerase chain reaction amplification from formalin-fixed and paraffin-embedded tissues.

To obtain an adequate quality and quantity of DNA from formalin-fixed and paraffin-embedded tissue, six different DNA extraction methods were compared. Four methods used deparaffinization by xylene followed by proteinase K digestion and phenol-chloroform extraction. The temperature of the different steps was changed to obtain higher yields and improved quality of extracted DNA. The remaining two methods used microwave heating for deparaffinization. The best DNA extraction method consisted of deparaffinization by microwave irradiation, protein digestion with proteinase K at 48 degrees C overnight, and no further purification steps. By this method, the highest DNA yield was obtained and the amplification of a 989-base pair beta-globin gene fragment was achieved. Furthermore, DNA extracted by means of this procedure from five gastric carcinomas was successfully used for single strand conformation polymorphism and direct sequencing assays of the beta-catenin gene. Because the microwave-based DNA extraction method presented here is simple, has a lower contamination risk, and results in a higher yield of DNA compared with the ordinary organic chemical reagent-based extraction method, it is considered applicable to various clinical and basic fields.

Semiquantitative immunoblot analysis of nm23-H1 and -H2 isoforms in adenocarcinomas of the lung: prognostic significance.

Total amounts of nm23 protein and relative levels of H1 and H2 isoforms were studied in 27 fresh-frozen samples of pulmonary adenocarcinoma and adjacent non-neoplastic tissues that were obtained at surgery. Semiquantitative immunoblotting with a monoclonal antibody (Pan-242) against nm23 protein demonstrated both isoforms, recognized as 20.5 kDa for H1 and 18.5 kDa for H2, to be present in all cases. Both H1 and H2 levels in neoplastic tissues were higher than in the corresponding non-neoplastic samples. Expression of H2 was usually greater than of H1. The H2/H1 ratio varied from 1.9 to 14.1 (mean value 5.2) in non-neoplastic tissues and 1.0-5.9 (mean value 2.5) in neoplastic tissues, although this ratio did not correlate with any prognostic factor like tumor size, nodal status or distant metastasis (TNM tumor stage). H1 and H2 levels were significantly lower (mean values 4.3 and 2.4) in well-differentiated than in moderately and poorly differentiated adenocarcinomas (8.3 and 3.0) (P < 0.03 and P < 0.05, respectively). These data indicate that H1 and H2 isoform levels correlate with histological differentiation, but not the metastatic potential or stage of pulmonary adenocarcinoma.

Chromogranin B (secretogranin I), a neuroendocrine-regulated secretory protein, is sorted to exocrine secretory granules in transgenic mice.

Chromogranin B (CgB, secretogranin I) is a secretory granule matrix protein expressed in a wide variety of endocrine cells and neurons. Here we generated transgenic mice expressing CgB under the control of the human cytomegalovirus promoter. Northern and immunoblot analyses, in situ hybridization and immunocytochemistry revealed that the exocrine pancreas was the tissue with the highest level of ectopic CgB expression. Upon subcellular fractionation of the exocrine pancreas, the distribution of CgB in the various fractions was indistinguishable from that of amylase, an endogenous constituent of zymogen granules. Immunogold electron microscopy of pancreatic acinar cells showed co-localization of CgB with zymogens in Golgi cisternae, condensing vacuoles/immature granules and mature zymogen granules; the ratio of immunoreactivity of CgB to zymogens being highest in condensing vacuoles/immature granules. CgB isolated from zymogen granules of the pancreas of the transgenic mice aggregated in a mildly acidic (pH 5.5) milieu in vitro, suggesting that low pH-induced aggregation contributed to the observed concentration of CgB in condensing vacuoles. Our results show that a neuroendocrine-regulated secretory protein can be sorted to exocrine secretory granules in vivo, and imply that a key feature of CgB sorting in the trans-Golgi network of neuroendocrine cells, i.e. its aggregation-mediated concentration in the course of immature secretory granule formation, also occurs in exocrine cells although secretory protein sorting in these cells is thought to occur largely in the course of secretory granule maturation.