Objective To investigate the effects and mechanism of papillary thyroid carcinoma susceptibility candidate 3(PTCSC3)on apoptosis of human neuroblastoma cells(SH-SY5Y)induced by glutamate(Glu).Methods SH-SY5Y cells were cultured in vitro and SH-SY5Y cells were injured by glutamate for 24 h.PTCSC3 was inhibited and overexpressed by transfection technique.Hoechst 33258 staining,flow cytometry,AO/EB staining and methylthiazolyl tetrazolium,MTT colorimetric assay were used to detect the effects of inhibition or overexpression of PTCSC3 on apoptosis and survival of cells in each group.Real-time fluorescence quantitative PCR(qPCR)was used to detect the gene expressions of PTCSC3,CCAAT/enhancer binding protein β(C/EBP-β)and Cbp/p300 binding transforming activator 4(Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 4)in cells of each group.Western blot was used to detect the protein expressions of C/EBP-β,brain-derived neurotrophic factor(BDNF),protein kinase B(Akt),B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and CITED4,Cyclin D1 and proto-oncogene n-Myc.Results 2 000 μmol·L-1 Glu could significantly damage PTCSC3 cells for 24 h.Compared with the model group,PTCSC3 inhibitor could effectively improve the increase of apoptosis rate and decrease of cell survival rate of SH-SY5Y cells induced by Glu,significantly reduce the expressions of BDNF-β gene and protein and Bax protein,promote the expressions of BDNF,p-Akt and Bcl-2 protein,and increase the expression levels of CITED4,Cyclin D1 and n-Myc.On the contrary,the results of overexpression of PTCSC3 were contrary to the above results.Conclusion PTCSC3 may improve the apoptosis and survival status of SH-SY5Y neurons induced by Glu by regulating C/EBP-β,and then regulating downstream signal pathways BDNF/Akt and CITED4/Cyclin D1.
维普
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