Vascular endothelial growth factor (VEGF) plays multifunctional roles in vascular permeability, repair and remodelling processes, in addition to the maintenance of vascular structure and function. In the present study, the potential of airway epithelial cell lines, BEAS-2B cells and A549 cells, to release and express VEGF in unstimulated and stimulated conditions was evaluated.The secretion and expression of VEGF were evaluated by enzyme-linked immunosorbant assay and by reverse transcriptase-polymerase chain reaction. The isoforms of released VEGF were determined by high-performance liquid chromatography.BEAS-2B cells and A549 cells released VEGF constitutively. Interleukin (IL)-1β and tumour necrosis factor (TNF)-α augmented the release of VEGF in a time- and dose-dependent manner. The released VEGF was 165 amino acid residues in either condition.Pseudomonas aeruginosalipopolysaccharide (LPS), interferon (IFN)-γ, smoke extract (SE), neutrophil elastase (NE), and bradykinin stimulated the release of VEGF. Keracinocyte growth factor (KGF), which reduces vascular permeability, also stimulated both cells to release VEGF. VEGF messenger ribonucleic acid (mRNA) was expressed both time- and dose-dependently at 2 h, and declined after 2 h in response to IL-1β and TNF-α. The expression of VEGF mRNA in airway epithelial cells was also augmented by LPS, IFN-γ, SE, NE, and KGF stimulation.These data suggest that airway epithelial cells may regulate the maintenance of vascular structure and function, as well as vascular permeability, repair and remodelling processes, in a variety of lung conditions by expressing vascular endothelial growth factor.

Purpose: Research has revealed abnormal activation of the hedgehog pathway in human malignancies.The present study was undertaken to examine the expression and functional involvement of the hedgehog pathway in endometrial tissues. Experimental Design: The expression of sonic hedgehog (Shh), patched (Ptch), Smoothened (Smo), and Gli1was examined in various endometrial tissues and endometrial carcinoma cell lines. The effect of hedgehog signaling on the proliferation of endometrial carcinoma cell lines was also examined. Results: The expression of Shh, Ptch, Smo, and Gli1was very weak in normal endometrium, but was increased in endometrial hyperplasia and carcinoma stepwisely with significant differences. There was no marked difference in the expression of these molecules in carcinomas according to stages and histologic grades. Treatment with cyclopamine, a specific inhibitor of the hedgehog pathway, for endometrial carcinoma Ishikawa and HHUA cells suppressed growth by 56% and 67%, respectively, compared with the control. The addition of recombinant Shh peptide to HHUA cells enhanced their proliferation by 41%. The silencing of Gli1using small interfering RNA (siGli1) resulted in the growth suppression and down-regulation of Ptch expression. In addition, the cyclopamine/siGli1-induced growth suppression was associated with the down-regulation of cyclins D1 and A and N-myc. No somatic mutations for ptch and smo genes were detected in the endometrial carcinoma cases examined. Conclusions:The abnormal activation of this pathway is involved in the proliferation of endometrial carcinoma cells possibly in an auto-/paracrine fashion, suggesting the possibility of the hedgehog pathway being a novel candidate for molecular targeting.

Purpose: Although several gene abnormalities have been reported in endometrial carcinoma, the genetic alterations have not fully been elucidated. Recent studies have revealed frequent activating mutations of the gene for BRAF, an effector of Ras protein in the mitogen-activated protein kinase pathway, in several malignancies. However, the prevalence and significance of BRAF mutations in endometrial carcinoma remain unclear. Experimental Design:We examined BRAF mutations in exons11and15 in 97 cases of endometrial carcinoma (endometrioid type, 78; nonendometrioid type,19), 9 cases of atypical endometrial hyperplasia, and 20 cases of normal endometrium by direct sequencing. In addition, mutations of KRAS and p53 and the immunohistochemical expression of hMLH1 and hMSH2 were also examined. Results: Of the 97 carcinomas and 9 hyperplasias, 20 (21%) and 1 (11%) had BRAF mutations, most of them at previously unreported sites. Twenty samples of normal endometrium and 21 samples of normal endometrium obtained from sites adjacent to neoplastic lesions had no BRAF mutations. There was no apparent difference in the prevalence of BRAF mutation among stages, histologic subtypes, or grades. Mutations of KRAS and p53 were found in 18 (19%) and 22 (23%) cases, and 65 (67%) and 92 (95%) cases showed positive immunostaining for hMLH1 and hMSH2, respectively. BRAF mutation was more frequently found in hMLH1-negative cases (12 of 32, 41%) than in hMLH1-positive cases (7 of 65, 11%; P = 0.008), suggesting that it is associated with an abnormal mismatch repair function. Conclusions: These findings suggest that mutations of the BRAF gene are partly involved in the malignant transformation of the endometrium.Endometrial carcinomas are now classified into two histologic subtypes: endometrioid type and nonendometrioid type (1); and the molecular genetic changes involved differ between the two (2 -4). In endometrioid-type carcinomas, mutations of PTEN, KRAS, b-catenin, and p53 genes and microsatellite instability have been reported in f5% to 50% of cases. By contrast, in nonendometrioid-type carcinomas, as exemplified by serous papillary adenocarcinoma, the p53 gene is reportedly mutated in f90% of cases. However, the genetic changes in endometrial carcinoma have not fully been elucidated.Raf proteins are cytoplasmic serine-threonine kinases and play central roles in the conserved Ras/Raf/mitogen-activated protein/extracellular signal-regulated kinase kinase/extracellular signal-regulated kinase pathway, acting to relay signals from activated Ras proteins via mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1/2 to p42/ 44 mitogen-activated protein kinase or extracellular signalregulated kinase 1/2, the key effector of the pathway (5, 6). A recent report has shown the presence of an activating mutation in one of three Raf protein subtypes, B-Raf, in human melanomas and colon cancers, and that the mutation of BRAF transforms NIH 3T3 cells independent of KRAS gene mutation (7). In addition, mutual exclusi...

Intraoperative ultrasonography can both safely and effectively localize pulmonary ground-glass opacities in a completely deflated lung. This procedure is also useful for the evaluation of surgical margins in a resected lung. Hence, ultrasonography may assist surgeons to perform minimally invasive lung resections with clear surgical margins during the treatment of solitary lung ground-glass opacity.