Project description:To characterize the mesothelial cell-mediated changes of gene expression in ovarian cancer cells, we performed RNA sequencing in OVCAR8 cells with and without mesothelial cell co-culture.
Project description:Asbestos-associated diseases remain a social burden worldwide. Our previous studies identified asbestos-induced iron-rich milieu for mesothelial cells with ceaseless macrophage ferroptosis. However, molecular mechanisms how this mutagenic milieu influences mesothelial cells have not been elucidated yet. Here, we propose a novel mechanism that extracellular vesicles (EVs) mediate asbestos-associated mutagenic factors to mesothelial cells. In a mice model of intraperitoneal crocidolite injection, mutagenic milieu highly expressed CD63, an exosomal marker. We then used a CD63-GFP labeled THP-1 macrophage model exposed to crocidolite/iron, which generated EVs under ferroptotic process. We observed that MeT-5A mesothelial cells can receive and internalize these EVs. Furthermore, we comprehensively analyzed the ferroptosis-dependent EVs (FedEVs) for transported proteins and identified ferritin heavy/light chains as major proteins. Therefore, we inferred that FedEVs transport iron from ferroptotic macrophages to mesothelial cells. RNA sequencing revealed that the mesothelial cells receiving higher amounts of the FedEVs were mitotic, especially at the S and G2/M phases, by the use of Fucci mesothelial cells. Nuclear 8-hydroxy-2`-deoxyguanosine and γ-H2AX were significantly increased in the recipient mesothelial cells after exposure to FedEVs. Collectively, we here demonstrate a novel mechanism that FedEVs act as a key mutagenic mediator by transporting iron, which contribute to asbestos-induced mesothelial carcinogenesis.
Project description:Characterization and Functional Analyses of Hepatic Mesothelial Cells in Mouse Liver Development. We used microarrays to detail the global programme of gene expression in mesothelial cells during mouse liver development. Mouse liver mesothelial cells were selected by a cell sorter at E12.5 and adult for RNA extraction and hybridization on Affymetrix microarrays. We obtained PCLP1+ cells and Mesothelin+ cells at each developmental stage in order to analyze expression profiles of mesothelial cells.
Project description:Characterization and Functional Analyses of Hepatic Mesothelial Cells in Mouse Liver Development. We used microarrays to detail the global programme of gene expression in mesothelial cells during mouse liver development.
Project description:Primary human omental mesothelial cells from multiple donors in a young (=normal) phase and senescent phase were treated with or without TGF-ß to induce fibrosis. Senescence was induced by up to 9 passages and confirmed by stopped cell division and positive senescence-associated β-galactosidase staining.
Project description:RNA-sequencing analysis was carried out on ascetic fluid-isolated mesothelial cells from ovarian cancer patients compared to control human peritoneal mesothelial cells to identify a mesothelial-mesenchymal gene signature.