Regulatory mechanism of IGSF1 mediated by miR-146a on invasion and metastasis of papillary thyroid carcinoma
Objective To investigate the regulation and mechanism of miR-146a on invasion and metastasis of TPC-1 cells.Methods CCK-8 and flow cytometry were used to detect cell activity;Transwell was used to detect cell invasion and apoptosis ability;scratch assay was used to detect cell healing ability;real-time fluorescence quantitative PCR was used to detect miR-146a and IGSF1 mRNA expression,and immunohis-tochemistry was used to detect IGSF1 expression.The protein expression of OPN and β-catenin was detec-ted by Western Blot,and the binding of miR-21 and IGSF1 was analyzed by dual luciferase reporter gene assay.Results The results of cell proliferation activity showed that the proliferation activity of TPC-1 cells(84.77±3.71)％was significantly reduced compared with of control group(100.00±0.00)％(P＜0.05).However,the cell proliferation activity of TPC-1 cells(119.20±5.81)％was significantly in-creased in miR-146a inhibitor group(P＜0.05).The flow cytometry results showed that the apoptosis rate of TPC-1 cells(20.33±1.26)％were increased significantly in the miR-146a mimics group compared with the control group(6.37±0.27)％,and the TPC-1 cell apoptosis rate(3.35±0.93)％was signifi-cantly decreased in the miR-146a inhibitor group(F=217.1,P＜0.05).The results of Transwell experi-ments and cell scratch experiment showed that the relative migration distance and the number of invading cells were decreased significantly at 24 h and 48 h(P＜0.05),while the relative migration distance and the number of invading cells in the miR-146a inhibitor group were increased significantly at 24 h and 48 h(P＜0.05).Compared with the control group,the miR-146a expression level was significantly increased;and the IGSF1 expression level was significantly decreased in the miR-146a mimics group(P＜0.05);and miR-146a expression level and IGSF1 expression level were significantly increased in miR-146a inhibitor group(P＜0.05).Compared with control group(0.92±0.09),β-catenin(0.54±0.08)and OPN(0.30 ±0.09)expression levels were all significantly reduced in the miR-146a mimics group(P＜0.05),andβ-catenin(1.26±0.07)and OPN(1.07±0.13)were significantly increased in miR-146a inhibitor group(P＜0.05).Dual-luciferase reporter gene assay showed that the results were statistically significant difference between wild type of miR-146a mimics group(0.64±0.11)and wild type of miR-146a mimics NC group(1.16±0.07)comparisons(P＜0.05).Conclusion miR-146a can interact with IGSF1 and regulate OPN and β-catenin.The expression level of catenin affects the progression of papillary thyroid carcinoma.
thyroid papillary carcinomamiRNAinvasionmechanism research
万方数据
维普
