微RNA-151-5p对甲状腺乳头状癌TPC-1细胞增殖和转移的影响及其机制

Effect of miR-151-5p on proliferation and metastasis of thyroid papillary carcinoma TPC-1 cells and its mechanism

目的探讨微RNA(miR)-151-5p对甲状腺乳头状癌TPC-1细胞增殖和转移的影响及其机制。方法将体外培养的TPC-1细胞分为Mimics-NC组(转染miR-151-5p mimics阴性对照)、Mimics组(转染miR-151-5p mimics)、Inhibitor-NC组(转染miR-151-5p inhibitor阴性对照)和Inhibitor组(转染miR-151-5p inhibitor),采用RT-PCR检测miR-151-5p表达,CCK-8法检测细胞增殖,Transwell小室检测细胞侵袭与迁移,免疫印迹检测细胞中E-cadherin、Vimentin、N-cadherin和SOX17蛋白的表达。采用双荧光素酶报告基因实验检测miR-151-5p与SOX17的靶向关系。结果与各自的非特性对照相比,转染miR-151-5p mimics后细胞中miR-151-5p和Vimentin、N-cadherin蛋白的表达水平明显升高,细胞增殖、侵袭和迁移能力明显增强,且细胞中E-cadherin、SOX17蛋白的表达水平明显降低(P<0.05);而转染miR-151-5p inhibitor后TPC-1细胞的变化与转染miR-151-5p mimics的结果正好相反。双荧光素酶报告基因实验证实SOX17是miR-151-5p的靶基因。结论miR-151-5p在甲状腺乳头状癌TPC-1细胞增殖、侵袭、迁移和上皮间质转化过程中发挥着重要的促进作用,其作用机制可能与靶向调控SOX17有关。

Objective To investigate the effect of miR-151-5p on proliferation and metastasis of thyroid papillary carcinoma TPC-1 cells and its mechanism.Methods TPC-1 cells cultured in vitro were divided into Mimics-NC group(transfected miR-151-5p mimics negative control),Mimics group(transfected miR-151-5p mimics),Inhibitor-NC group(transfected miR-151-5p inhibitor negative control)and Inhibitor group(transfected miR-151-5p inhibitor).The expression of miR-151-5p was detected by RT-PCR,cell proliferation by CCK-8 assay,cell invasion and migration by Transwell chamber assay,and protein expression of E-cadherin,Vimentin,N-cadherin and SOX17 by Western blotting.The targeting relationship between miR-151-5p and SOX17 was detected by dual luciferase reporter gene assay.Results Compared with the respective non-characterized controls,cells transfected with miR-151-5p mimics showed significantly higher expression levels of miR-151-5p,Vimentin and N-cadherin proteins,promoted cell proliferation,invasion and migration,and suppressed expression of E-cadherin and SOX17 proteins(P<0.05).These changes were exactly opposite to those found in TPC-1 cells transfected with miR-151-5p inhibitor.Dual luciferase reporter gene experiments confirmed that SOX17 is a target gene of miR-151-5p.Conclusion miR-151-5p plays an important promoting role in the proliferation,invasion,migration and epithelial-mesenchymal transition of papillary thyroid carcinoma TPC-1 cells,and its mechanism may be related to targeted regulation of SOX17.