摘要
目的探讨miR-197-3p是否通过靶向调控恶性脑瘤缺失1基因(DMBT1)影响甲状腺癌细胞增殖、迁移和侵袭。方法RT-qPCR检测健康人甲状腺细胞Nthy-ori 3-1和甲状腺癌细胞SW579、CGTHW-1中miR-197-3p表达;MTT法检测SW579细胞增殖;Transwell小室法检测SW579细胞迁移和侵袭;双荧光素酶报告基因实验验证miR-197-3p是否靶向DMBT1;Western blot检测细胞DMBT1、cyclinD1、p21、MMP-2和E-cadherin蛋白表达。结果与Nthy-ori 3-1细胞比较,SW579和CGTHW-1细胞中miR-197-3p相对表达量升高(P<0.05);抑制miR-197-3p表达后,SW579细胞的增殖、迁移和侵袭能力明显受到抑制,细胞中cyclinD1蛋白和MMP-2蛋白表达降低而p21蛋白和E-cadherin蛋白表达升高;SW579细胞中miR-197-3p靶向负调控DMBT1的表达;过表达DMBT1明显抑制SW579细胞增殖、迁移和侵袭,而抑制DMBT1则能逆转miR-197-3p对SW579细胞增殖、迁移和侵袭的影响。结论miR-197-3p通过靶向调控DMBT1的表达,抑制甲状腺癌细胞增殖、迁移和侵袭。
Objective To investigate whether miR-197-3p affects the proliferation,migration and invasion of thyroid cancer cells through regulation of deletion of malignant brain tumors 1(DMBT1)expression.MethodsThe expression of miR-197-3p in human normal thyroid cells Nthy-ori 3-1 and thyroid cancer cell lines SW579 and CGTHW-1 was detected by RT-qPCR,MTT assay was used to detect the proliferation of SW579 cells,and Transwell chamber assay was used to detect the migration and invasion of SW579 cells,the dual luciferase reporter gene assay verified whether miR-197-3p targets DMBT1,and the expression of DMBT1,cyclin D1,p21,MMP-2and E-cadherin in the cells was detected by Western blot.Results Compared with Nthy-ori 3-1 cells,the relative expression of miR-197-3p was increased in SW579 and CGTHW-1 cells(P<0.05).After inhibiting the expression of miR-197-3p,the proliferation,migration and invasion of SW579 cells were significantly inhibited,and the expression of cyclin D1 and MMP-2 proteins was decreased and the expression of p21 and E-cadherinproteins was increased.miR-197-3p targeting negatively regulates DMBT1 expression in SW579 cells.Over-expression of DMBT1 significantly inhibited the proliferation,migration and invasion of SW579 cells,while inhibition of DMBT1 reversed the effect of miR-197-3p on proliferation,migration and invasion of SW579 cells.Conclusions miR-197-3p inhibits the proliferation,migration and invasion of thyroid cancer cell regulation of DMBT1 expression.
作者
曹邓晗
陈奎生
CAO Deng-han;CHEN Kui-sheng(Department of Endocrinology, Zhengzhou First People’s Hospital, Zhengzhou 450004;Department of Pathology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052,China)
出处
《基础医学与临床》
CSCD
2020年第5期644-650,共7页
Basic and Clinical Medicine
基金
河南省医学科技攻关计划项目(201503221)。