| Objectives To investigate the effect of miR-454 on the proliferation,migration and invasion of papillary thyroid cancer cells.Methods Normalized culture of the papillary thyroid cancer cell line BCPAP,transfection of cells with CY3-labeled fluorescent control and lipofectamine 2000.Observe the number of cells glowing under fluorescence microscopy and the total number of cells in the same field of view to determine transfection efficiency.Using lipofectamine 2000,respectively transfect PBS solution,miR-454 inhibitor,inhibitor control,miR-454 mimic and mimic control,marked as blank group,inhibitor group,inhibitor control group,mimic group and mimic control group.Quantitative real-time PCR(q-PCR)was used to detect the expression level of miR-454 after transfection.Cell proliferation capacity was detected by CCK-8 cell proliferation experiments.Cell migration capacity was detected by single-layer cell scratch injury repair experiments.Cell invasion ability was detected by Transwell invasion experiments.Expression of RPRD1 A and β-catenin proteins was detected by using western blotting experiments.Results 1 After transfection of CY3-labeled fluorescent control to intracellular 24 h,the transfection efficiency can be seen under fluorescence microscopy of more than 60%.2The q-PCR results showed that the relative expression of miR-454 in the inhibitor group was significantly reduced compared with the inhibitor control group(P<0.05).And the relative expression of miR-454 in the mimic group was significantly increased compared with the mimic control group(P<0.05).3 The results of CCK-8 cell proliferation experiments showed that the optical density value(OD value)of the inhibitor group was significantly reduced compared with the inhibitor control group in the 24 h,48h,72 h and96h time periods(P<0.05).And the OD value of the mimic group was significantly higher than that of the mimic control group(P<0.05).4 The results of the single-layer cell scratch injury repair experiment showed that the 48-hour scratch healing rate of the inhibitor group was significantly reduced compared with the inhibitor control group(P<0.05).The 48-hour scratch healing rate of the mimic group was significantly higher than that of the mimic control group(P<0.05).5 The results of Transwell invasion experiment showed that the number of cells crossing the chamber in the inhibitor group was significantly reduced compared with the inhibitor control group(P<0.05).And the number of cells passing through the chamber in the mimic group was significantly higher than that in the mimic control group(P<0.05).6 The results of the western blotting experiment showed that the expression level of RPRD1 A protein in the inhibitor group was significantly increased,and the expression level of β-catenin protein was significantly decreased compared with the inhibitor control group(P<0.05);the expression level of RPRD1 A protein in the mimic group decreased significantly compared with the mimic control group,while the expression of β-catenin protein increased significantly(P<0.05).Conclusions 1 miR-454 promotes the proliferation,migration,and invasion of papillary thyroid cancer cells.2 miR-454 may regulate the Wnt/β-catenin signaling pathway via RPRD1 A.3 miR-454 is promising to provide new ideas for the future clinical treatment of papillary thyroid cancer.Figure 11;Table 2;Reference 142... |
