Molecular mechanism of P75 NTR gene-induced apoptosis in tongue squamous cell carcinoma cell
Author(s): Zhang Zhaotao, Zhang Fenghe, Tong Dongdong, Li Qing, Wang Jinbing, Zhang Xinlian
Pages: 481-485
Year: 2016 Issue: 7
Journal: Journal of International Oncology
Keyword: Apoptosis; Nerver growth factor; NF-Kappa B;
Abstract: Objective To investigate the molecular mechanism of P75NTR gene-induced apoptosis in tongue squamous cell carcinoma Tca8113 cell lineage.Methods P75NTR specific siRNA was transferred into P75NTR positive tongue squamous cell carcinoma Tca8113 cells.P75NTR positive Tca8113 cells were divided into 4 groups:blank group (without transfection),negative control group (transfected with negative control siRNA ), experiment group-776 (transfected with siRNA-P75NTR-776 ) and experiment group-1234 (transfected with siRNA-P75NTR-1234).Transfection efficiency and cell apoptosis were detected by flow cytometry.The interference effect of P75NTR mRNA expression was detected by fluorescence quantitative PCR. 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide assay was applied in measuring cell prolife-ration.The protein changes of P75NTR were detected by Western blotting.The distributions of nuclear factor-κB(NF-κB)of cells were observed by cell immunofluorescence labeling method.Results The transfection efficiency was 30%.The apoptosis rate of experiment group-776,experiment group-1234 and negative control group was (20.35 ±0.18)%,(12.32 ±1.51)% and (2.63 ±0.10)% respectively.Compared with the negative control group,the differences of the former two group had statistical significance (t =177.20,P <0.005;t =37.12,P <0.005).The P75NTR gene interference was successful.The inhibition rate of P75NTR protein reached 31% in experiment group-776.The cell viability of Tca8113 cells after P75NTR-siRNA inter-
ference was 70.02%,78.01% and 95.81% in experiment group-776,experiment group-1234 and negative control group.And there were significant differences between experiment group-776 and negative control group (χ2 =235.3,P <0.010),and between experiment group-1234 and negative control group (χ2 =117.5,P <0.005 ).NF-κB distribution was increased in cell cytoplasm in the interference group than that in control group.Conclusion P75NTR may promote the proliferation or inhibit the apoptosis of tongue squamous cell carcino-ma,and the molecular mechanism may be correlated with hindering the transportion of NF-κB into cell nuclear.
Pages: 481-485
Year: 2016 Issue: 7
Journal: Journal of International Oncology
Keyword: Apoptosis; Nerver growth factor; NF-Kappa B;
Abstract: Objective To investigate the molecular mechanism of P75NTR gene-induced apoptosis in tongue squamous cell carcinoma Tca8113 cell lineage.Methods P75NTR specific siRNA was transferred into P75NTR positive tongue squamous cell carcinoma Tca8113 cells.P75NTR positive Tca8113 cells were divided into 4 groups:blank group (without transfection),negative control group (transfected with negative control siRNA ), experiment group-776 (transfected with siRNA-P75NTR-776 ) and experiment group-1234 (transfected with siRNA-P75NTR-1234).Transfection efficiency and cell apoptosis were detected by flow cytometry.The interference effect of P75NTR mRNA expression was detected by fluorescence quantitative PCR. 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide assay was applied in measuring cell prolife-ration.The protein changes of P75NTR were detected by Western blotting.The distributions of nuclear factor-κB(NF-κB)of cells were observed by cell immunofluorescence labeling method.Results The transfection efficiency was 30%.The apoptosis rate of experiment group-776,experiment group-1234 and negative control group was (20.35 ±0.18)%,(12.32 ±1.51)% and (2.63 ±0.10)% respectively.Compared with the negative control group,the differences of the former two group had statistical significance (t =177.20,P <0.005;t =37.12,P <0.005).The P75NTR gene interference was successful.The inhibition rate of P75NTR protein reached 31% in experiment group-776.The cell viability of Tca8113 cells after P75NTR-siRNA inter-
ference was 70.02%,78.01% and 95.81% in experiment group-776,experiment group-1234 and negative control group.And there were significant differences between experiment group-776 and negative control group (χ2 =235.3,P <0.010),and between experiment group-1234 and negative control group (χ2 =117.5,P <0.005 ).NF-κB distribution was increased in cell cytoplasm in the interference group than that in control group.Conclusion P75NTR may promote the proliferation or inhibit the apoptosis of tongue squamous cell carcino-ma,and the molecular mechanism may be correlated with hindering the transportion of NF-κB into cell nuclear.
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