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Effects of lentivirus-mediated RWDD3 silencing on proliferation and invasion of human glioma U251 cells
Authors:FAN Yang-hua  ZHU Xin-gen  L&#  Shi-gang  WU Miao-jing  CHAI Yi  YE Min-hua  XIAO Bing  WU Lei
Institution:Department of Neurosurgery, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China
Abstract:AIM: To investigate the effect of RWDD3 gene silencing on the biological characteristics of human glioma U251 cells.METHODS: A lentiviral vector expressing RWDD3 shRNA was constructed and transfeeted into the U251 cells. The expression of RWDD3 at mRNA and protein levels was detected by real-time PCR and Western blot, respectively. The cell activity was determined by MTT assay. The colony formation ability was detected by the colony formation assay. The cell proliferation ability was detected by BrdU incorporation assay. The cell invasion and migration were evaluated by Transwell assay. Flow cytometry was used to monitor the changes of cell cycle distribution and apoptosis.RESULTS: Recombinant lentivirus was successfully transfected into U251 cells. Compared with the cells transfected with the scrambled shRNA and control cells, the cell activity, colony formation ability, and the invasive and migratory activities were inhibited, the cell cycle was arrested in G0/G1 phase, and the apoptosis was increased in the U251 cells transfected with RWDD3 shRNA(P<0.05).CONCLUSION: RWDD3 plays a vital role in proliferation and invasion of glioma cells. It may serve as a potential target of gene therapy for glioma.
Keywords:Glioma  RWDD3  Lentivirus  Proliferation  Invasion  
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