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- W2384004249 abstract "Objective This study was to design a new recombinant plasmid expressing the carboxylic terminal peptide of DAPK (DCTP) to inhibit SH-SY5Y cells apoptosis induced by glutamate acid taking DAPK as the target.The mechanism of inhibition of DAPK by DCTP was explored.Methods The DCTP coded sequence was amplified by PCR with the cDNA of DAPK as template,and oriented to insert into the fusion expression plasmid pEGFPN1.The recombinant plasmid was transfected into SH-SY5Y cells.DCTP expression was detected with RT-PCR.The stably expressed DCTP cell line was established after the cells were screened by G418.MTT method,Hoechst33258-staining and flow cytometry were used to measure cell survival,apoptosis and necrosis after SH-SY5Y cells were treated with glutamate for 24h.The changes in DAPK expression and DAPK phosphorylation were deteced by Western bloting.Results Glutamate inhibited the growth of SH-SY5Y cells in a dose-dependent manner.The inhibition rate of 40mmol/L glutamic acid was 51.7%,the apoptotic rate and necrosis rate were 26.1% and 27.7% in the presence of 40mmol/L glutamate,respectively,while they were respectively 0.08% and 0% in control group.The phosphorylation levels of DAPK descreased in the presence of glutamate.Observation with fluorescence microscopy revealed that DCTP expression products were located in cytoplasm.When the cells of transfected DCTP and control cells were treated with 40mmol/L of glutamate for 24h,the apoptotic ratio of DCTP-transfected cell was 43.7%,while of control cell was 62.2%,as mearsured by flow cytometry.Compared with the vector-transfected control cells induced by glutamate,the expression of DAPK and the phosphorylation level of DAPK were not significantly changed in DCTP-transfected cells induced by glutamate.Conclusions It is verified that the DCTP expression products are located in the cytoplasm of SH-SY5Y cells.It can prevent apoptosis of SH-SY5Y cells induced by glutamate without affecting the expression and the phosphorylation level of DAPK." @default.
- W2384004249 created "2016-06-24" @default.
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- W2384004249 date "2010-01-01" @default.
- W2384004249 modified "2023-09-22" @default.
- W2384004249 title "Inhibition of glutamate induced apoptosis of SH-SY5Y cells by C-terminal tail of DAPK" @default.
- W2384004249 hasPublicationYear "2010" @default.
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