过表达miR-23b-3p对甲状腺未分化癌FRO细胞侵袭和迁移的影响

目的探讨miR-23b-3p对甲状腺未分化癌(ATC)FRO细胞侵袭和迁移能力的影响。 方法qRT-PCR法检测52例ATC癌组织及其癌旁组织中miR-23b-3p和锌指E盒结合同源框1(ZEB1)的表达水平,并分析两者的相关性。培养人ATC细胞株FRO,将miR-23b-3p mimic及其阴性对照转染至FRO细胞中,qRT-PCR检测miR-23b-3p和ZEB1 mRNA表达水平。体外三维培养观察细胞血管生成拟态(VM)形成能力；Transwell法检测细胞侵袭和迁移能力；Western blotting检测ZEB1、血管内皮生长因子(VEGF)、基质金属蛋白酶(MMP)-2和MMP-9蛋白表达水平,双荧光素酶报告实验验证miR-23b-3p和ZEB1的靶向调控关系。 结果ATC组织中miR-23b-3p表达水平低于癌旁组织,而ZEB1的表达水平高于癌旁组织,且两者在ATC组织中的表达水平呈负相关。过表达miR-23b-3p能显著下调ZEB1表达水平,抑制FRO细胞VM形成以及侵袭和迁移能力,同时抑制细胞中VEGF、MMP-2和MMP-9蛋白表达。荧光素酶报告实验证实ZEB1是miR-23b-3p的靶基因。 结论过表达miR-23b-3p可能通过靶向下调ZEB1表达抑制FRO细胞的侵袭和迁移能力。

Effects of overexpression of miR-23b-3p on the invasive and migratory capacity of thyroid undifferentiated FRO cells

AimTo investigate the effects of overexpression of miR-23b-3p on the ability of invasion and migration of anaplastic thyroid cancer FRO cells. MethodsqRT-PCR was used to detect the expression levels of miR-23b-3p and zinc finger E-box-binding homology box 1(ZEB1) in cancer tissues and adjacent tissues of 52 ATC patients, and the correlation between them was analyzed. Human ATC cell line FRO was cultured, and miR-23b-3p mimic and its negative control were transfected into FRO cells. The expression levels of miR-23b-3p and ZEB1 mRNA were detected by qRT-PCR. The ability of vasculogenic mimicry (VM) formation was observed by three-dimensional culture in vitro. Transwell assay was used to detect cell invasion and migration. Western blotting was used to detect the protein expression levels of ZEB1, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2 and MMP-9. Dual luciferase reporter assay was used to verify the targeted regulatory relationship between miR-23b-3p and ZEB1. ResultsThe expression level of miR-23b-3p in ATC tissues was significantly lower than that in the corresponding adjacent normal tissues, while the expression level of ZEB1 was significantly increased, there was a negative correlation between their expression in ATC tissues. Overexpression of miR-23b-3p could significantly down-regulate the mRNA and protein expression levels of ZEB1, inhibit the invasion and migration, as well as the VM formation of FRO cells, and inhibit proteins the expression of VEGF, MMP-2 and MMP-9. ConclusionOverexpression of miR-23b-3p may inhibit the invasion and metastasis ability of FRO cells by targeting down-regulation of ZEB1 expression.