title: Downregulation of B-cell lymphoma/leukemia-2 by overexpressed microRNA 34a enhanced titanium dioxide nanoparticle-induced autophagy in BEAS-2B cells.
abstract: Titanium dioxide (TiO2) nanoparticles (TNPs) are manufactured worldwide for a wide range of applications and the toxic effect of TNPs on biological systems is gaining attention. Autophagy is recognized as an emerging toxicity mechanism triggered by nanomaterials. MicroRNA 34a (miR34a) acts as a tumor suppressor gene by targeting many oncogenes, but how it affects autophagy induced by TNPs is not completely understood. Here, we observed the activation of TNP-induced autophagy through monodansylcadaverine staining and LC3-I/LC3-II conversion. Meanwhile, the transmission electron microscope ultrastructural analysis showed typical morphological characteristics in autophagy process. We detected the expression of miR34a and B-cell lymphoma/leukemia-2 (Bcl-2). In addition, the underlying mechanism of TNP-induced autophagy was performed using overexpression of miR34a by lentivirus vector transfection. Results showed that TNPs induced autophagy generation evidently. Typical morphological changes in the process of autophagy were observed by the transmission electron microscope ultrastructural analysis and LC3-I/LC3-II conversion increased significantly in TNP-treated cells. Meanwhile, TNPs induced the downregulation of miR34a and increased the expression of Bcl-2. Furthermore, overexpressed miR34a decreased the expression of Bcl-2 both in messenger RNA and protein level, following which the level of autophagy and cell death rate increased after the transfected cells were incubated with TNPs for 24 hours. These findings provide the first evidence that overexpressed miR34a enhanced TNP-induced autophagy and cell death through targeted downregulation of Bcl-2 in BEAS-2B cells.
authors.affiliation: Department of Occupational Health and Environmental Health, School of Public Health, Capital Medical University, Beijing, People's Republic of China; Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Wenlin
authors.initials: W
authors.lastname: Bai
authors.affiliation: Department of Occupational Health and Environmental Health, School of Public Health, Capital Medical University, Beijing, People's Republic of China; Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Yujiao
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Occupational Health and Environmental Health, School of Public Health, Capital Medical University, Beijing, People's Republic of China; Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Pengling
authors.initials: P
authors.lastname: Sun
authors.affiliation: Department of Occupational Health and Environmental Health, School of Public Health, Capital Medical University, Beijing, People's Republic of China; Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing, People's Republic of China.
authors.forename: Ai
authors.initials: A
authors.lastname: Gao
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects*; D001980:Bronchi / Q000166:cytology; D015536:Down-Regulation / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D035683:MicroRNAs / Q000187:drug effects / Q000235:genetics*; D046529:Microscopy, Electron, Transmission; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism; D014025:Titanium / Q000633:toxicity*; D014162:Transfection
chemical_list: C477012:MAP1LC3A protein, human; C522425:MIRN34 microRNA, human; D035683:MicroRNAs; D008869:Microtubule-Associated Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; C009495:titanium dioxide; D014025:Titanium
keywords: Bcl-2; autophagy; cell death; cytotoxicity; lentivirus; miR34a; titanium dioxide nanoparticles