Effect Of FOXQ1-siRNA On EMT Of Thyroid Carcinoma TPC-1 Cells

Objectives To investigate the effect of forkhead box Q1?Forkhead box Q 1,FOXQ1?-small interfering RNA?siRAN?on epithelial-mesenchymal transition?EMT?of thyroid papillary carcinoma TPC-1 cells and its mechanism.Methods The expression of FOXQ1 protein in normal thyroid Nthy-ori3-1 cells and thyroid papillary carcinoma TPC-1 cells was detected by Western blot.The synthetic nonsense siRNA?NC-siRNA?and the three-entry siRNA?FOXQ1-siRNA?were transfected into TPC-1 cells of the thyroid papillary carcinoma by transfection reagent lipofectamine^{T M}2000.The transfection efficiency was evaluated by observing the expression of green fluorescent protein in the cells by fluorescence microscopy.Real-time quantitative PCR?qRT-PCR?and Western blot were used to detect the expression of FOXQ1 when the interference efficiency reac hed 85%or more.After screening the best interference sequences,the experiments were divided into three groups:control group plus equal amount of phosphate buffer?PBS??negative control group?NC-siRNA group??transfection group?FOXQ1-3 group?.MTT assay was used to detect the changes of cell proliferation activity at each time point?24 h,48 h,72 h,96 h?.Tumor cell isolation and adhesion assays detect changes in cell adhesion in vitro.Transwell invasion and migration assays were used to examine the effects of FOXQ1-siRNA on cell invasion and migration in vitro.Western blot was used to detect expression of FOXQ1,protein kinase B?AKT?,phosphorylated protein kinase B?p-AKT?,E-cadherin,N-cadherin and Vimentin.Results Western blot results showed that the expression levels of FOXQ1 protein in human thyroid normal cells Nthy-ori3-1 and thyroid papillary carcinoma TPC-1 cells were:?0.52±0.06???0.91±0.05?;papillary thyroid carcinoma TPC-1 The expression of FOXQ1protein in cells was significantly increased?P<0.05?.The results of qRT-PCR and Western blot showed that the three transfection sequences had interference effects on the FOXQ1protein of TPC-1 cells,and the FOXQ1-3 group had the best effect?P<0.05?,and the effective interference sequences were successfully screened.The results of MTT showed that compared with control group and NC-siRNA group,FOXQ1-3 group had significant inhibitory effects on cell proliferation at 24h,48h,72h and 96h?P<0.001?.The results of tumor cell adhesion assay showed that the separation ability and heterologous adhesion ability of FOXQ1-3 group cells were lower than that of control group and NC-siRNA group?P<0.05?.The results of Transwell invasion and migration assay showed that the invasion and migration ability of FOXQ1-3 group cells were significantly inhibited compared with control group and NC-siRNA group?P<0.001?.Western blot analysis showed that after FOXQ1-siRNA transfected cells,the expression of AKT protein in FOXQ1-3 group was unchanged,the expression of E-cadherin protein was increased,and the expression levels of FOXQ1,p-AKT,N-cadherin and Vimentin were increased.Significantly reduced?P<0.001?.Conclusions Silencing the FOXQ1 gene inhibits the invasion and migration of human papillary thyroid carcinoma TPC-1 cells by reversing EMT,which may be achieved through the PI3K/AKT signaling pathway.Confirmed that FOXQ1 gene plays a crucia l role in PTC treatment and prognosis.Figure 8;Table 6;Reference 120...