Glioblastoma (GBM) is the most common brain tumor in adults and the mesenchymal GBM subtype was reported to be the most malignant, presenting severe hypoxia and necrosis. Here, we investigated the possible role of a hypoxic microenvironment for inducing a mesenchymal and invasive phenotype. The exposure of non-mesenchymal SNB75 and U87 cells to hypoxia induced a strong change in cell morphology that was accompanied by enhanced invasive capacity and the acquisition of mesenchymal marker expression. Further analyses showed the induction of HIF1α and HIF2α by hypoxia and exposure to digoxin, a cardiac glycoside known to inhibit HIF1/2 expression, was able to prevent hypoxia-induced mesenchymal transition. ShRNA-mediated knockdown of HIF1α, and not HIF2α, prevented this transition, as well as the knockdown of the EMT transcription factor ZEB1. We provide further evidence for a hypoxia-induced mesenchymal shift in GBM primary material by showing co-localization of GLUT1, ZEB1 and the mesenchymal marker YKL40 in hypoxic regions of the tumor. Collectively, our results identify a HIF1α-ZEB1 signaling axis that promotes hypoxia induced mesenchymal shift and invasion in GBM in a cell line dependent fashion.
This work presents the cross-sections for radioactive nuclide production in 56 Fe(p,x) reactions determined in six experiments using 300, 500, 750, 1000, 1500, and 2600 MeV protons of the external beam from the ITEP U-10 proton accelerator. In total, 221 independent and cumulative yields of radioactive residuals of half-lives from 6.6 min to 312 days have been obtained. The radioactive product nuclide yields were determined by direct g-spectrometry. The measured data have been compared with the experimental data obtained elsewhere by the direct and inverse kinematics methods and with calculation results of 15 different codes that simulated hadron-nucleus interactions: MCNPX (INCL, CEM2k, BERTINI, ISABEL), LAHET (BERTINI, ISABEL), CEM03 (.01, .G1, .S1), LAQGSM03 (.01, .G1, .S1), CASCADE-2004, LAHETO, and BRIEFF. Most of the data obtained here are in a good agreement with the inverse kinematics results and disprove the results of some earlier activation measurements that were quite different from the inverse kinematics measurements. The most significant calculation-to-experiment differences are observed in the yields of the A<30 light nuclei, indicating that further improvements in nuclear reaction models are needed, and pointing out as well to a necessity of more complete experimental measurements of such reaction products.
Understanding the molecular and cellular processes underlying the development, maintenance, and progression of Barrett’s esophagus (BE) presents an empirical challenge because there are no simple animal models and standard 2D cell culture can distort cellular processes. Here we describe a three-dimensional (3D) cell culture system to study Barrett’s esophagus. BE cell lines (CP-A, CP-B, CP-C, and CP-D) and esophageal squamous keratinocytes (EPC2) were cultured on a matrix consisting of esophageal fibroblasts and collagen. Comparison of growth and cytokeratin expression in the presence of all-trans retinoic acid or hydrochloric acid was made by immunohistochemistry and Alcian Blue staining to determine which treatments produced a BE phenotype of columnar cytokeratin expression in 3D culture. All-trans retinoic acid differentially affected the growth of BE cell lines in 3D culture. Notably, the non-dyplastic metaplasia-derived cell line (CP-A) expressed reduced squamous cytokeratins and enhanced columnar cytokeratins upon ATRA treatment. ATRA altered the EPC2 squamous cytokeratin profile towards a more columnar expression pattern. Cell lines derived from patients with high grade dysplasia already expressed columnar cytokeratins and therefore did not show a systematic shift toward a more columnar phenotype with ATRA treatment. ATRA treatment however did reduce the squamoid-like multilayer stratification observed in all cell lines. As the first study to demonstrate long term 3D growth of BE cell lines, we have determined that BE cells can be cultured for at least three weeks on a fibroblast/collagen matrix and that the use of ATRA causes a general reduction in squamous-like multilayered growth and an increase in columnar phenotype with the specific effects cell-line dependent.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.