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Objective:To analyze the relationship between DNA methyltransferase 1 (DNMT1) and forkhead box O3a (FOXO3a) in colon cancer and the diagnostic efficacy of combined detection in predicting the occurrence of colon cancer by detecting the levels of DNMT1 and FOXO3a in serum of colon cancer patients.Methods:A total of 105 patients with colon cancer diagnosed and treated in Xi′an International Medical Center Hospital from September 2019 to September 2020 were selected as the colon cancer group, and 65 patients with colon polyps diagnosed by biopsy during the same period were selected as control group. The levels of DNMT1 and FOXO3a in serum of patients were detected by real-time fluorescence quantitative PCR. Pearson correlation coefficient method was used to analyze the correlation between the levels of DNMT1 and FOXO3a in serum of patients with colon cancer. Subject operating characteristic curve was used to evaluate the diagnostic values of DNMT1 and FOXO3a levels in colon cancer.Results:The serum levels of DNMT1 in the control group and colon cancer group were 0.93±0.28 and 1.34±0.35, compared with the control group, the level of DNMT1 in the colon cancer group was significantly higher, with a statistically significant difference ( t=7.990, P<0.001). The serum levels of FOXO3a were 1.04±0.39 and 0.69±0.18, compared with the control group, the level of FOXO3a in the colon cancer group was significantly lower, with a statistically significant difference ( t=7.940, P=0.001). The serum levels of DNMT1 and FOXO3a in patients with colon cancer were negatively correlated ( r=-0.687, P<0.001). The area under the curve (AUC) of DNMT1 predicting colon cancer was 0.843, the sensitivity was 71.40%, and the specificity was 90.80%. The AUC of FOXO3a predicting colon cancer was 0.812, the sensitivity was 88.60%, and the specificity was 67.70%. The AUC of the two combined predicting colon cancer was 0.859, the sensitivity was 89.50%, and the specificity was 92.30%. Compared with FOXO3a single detection, the predictive value of combined detection of DNMT1 and FOXO3a were higher ( Z=1.982, P=0.047). Conclusion:The level of DNMT1 in the serum of patients with colon cancer is increased, while the level of FOXO3a is decreased. There is a negative correlation between them in the serum of patients with colon cancer. The combined detection of the DNMT1 and FOXO3a can effectively improve the diagnostic value of colon cancer.
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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.
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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.
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<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>