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 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.
ABSTRACT
Objective:To study the effect of p75NTR gene on the apoptosis of tongue squamous cell carcinoma Tca8113 cells. Methods:p75NTR +Tca8113 cells were isolated from Tca 8113 cell line and transfected by p75NTR siRNA using lipofectamine. p75NTR mRNA and protein expression was examined by real time RT-PCR and western blot respectively.Cell proliferation was stud-ied by MTT assay,cell apoptosis was examined by flow cytometry.Results:Proliferation of p75NTR + cells was faster than that of p75NTR - cells.Transfection of p75NTR siRNA inhibited p75NTR mRNA and protein expression in p75NTR + Tca8113 cells,inhibi-ted the proliferation and increased the apoptosis of the cells.Conclusion:p75NTR gene plays a role in the apoptosis of Tca8113 cells.
ABSTRACT
<p><b>OBJECTIVE</b>To study the biological characteristics of p75 neurotrophin receptor positive (p75(NTR+)) tongue squamous cell carcinoma cells which were separated by flow cytometry cell sorting.</p><p><b>METHODS</b>To determine the biological characteristics of p75(NTR+) cells which were separated from Tca-8113 and Cal-27 tongue squamous cell carcinoma cells by flow cytometry cell sorting, including study the capacity of cloning, 3-(4,5)-demethylthiazo(z-y1)-3,5-diphenytetrazoliumromide (MTT) assay, wound healing assay. p75(NTR+) cells with non-sorted cells were as control group.</p><p><b>RESULTS</b>In Tca-8113 and Cal-27 tongue squamous cell carcinoma cell lines, the percentage of p75(NTR+) cells were 3.1% and 1.9%. Compared with p75(NTR+) cells with non-sorted cells, p75(NTR+) cells possess higher capacity of cloning (Tca-8113, P=0.024; Cal-27, P=0.009). The percentage of p75(NTR+) cells of the progeny cells generated from monoclonal p75(NTR+) cells decreased to 14.5% (Tca-8113) and 5.8% (Cal-27) after cultured two weeks. p75(NTR+) cells possessed higher proliferation ability and higher metastasis ability than non-sorted cells.</p><p><b>CONCLUSION</b>p75(NTR+) cells isolated from tongue squamous cell carcinoma have the characteristics of cancer stem cells.</p>