Department of Obstetrics and Gynecology, Dafeng Hospital of Traditional Chinese Medicine, Yancheng, Jiangsu, China.
Crit Rev Eukaryot Gene Expr. 2021;31(2):63-73. doi: 10.1615/CritRevEukaryotGeneExpr.2021037776.
The purpose of this study was to elucidate the role that the miR-18a-5p/THBD regulatory pathway plays in endometrial cancer (EC), which could provide a theoretical basis for potential therapeutic targets. Differentially expressed genes in EC tissue and normal tissue were determined by bioinformatics analysis. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to compare the expression of miR-18a-5p and THBD mRNA in normal human endometrial cells and human EC cells. CCK-8 assay was used to compare the proliferative ability of EC cells in different treatment groups. Transwell assay was used to detect the migratory and invasive abilities of EC cells in different treatment groups. Dual-luciferase assay was used to verify the targeting relationship between miR-18a-5p and THBD. Western blot assay was used to detect THBD protein expression level. qRT-PCR results showed that miR-18a-5p was significantly upregulated in EC cells, and expression of its target gene, THBD, was significantly downregulated. CCK-8 and transwell assays showed that miR-18a-5p could enhance the proliferative, migratory, and invasive abilities of EC cells, whereas THBD could weaken those abilities. Dual-luciferase assay confirmed that miR-18a-5p could negatively regulate THBD expression. In addition, rescue experiments revealed that the oncogenic effect of miR-18a-5p on EC cells was inhibited by THBD overexpression. We conclude that miR-18a-5p could promote the proliferation, migration, and invasion of EC cells by targeting and downregulating THBD expression, and the miR-18a-5p/THBD regulatory pathway might be a therapeutic target. The results of this study may serve as a theoretical basis for related drug development.
本研究旨在阐明 miR-18a-5p/THBD 调控通路在子宫内膜癌（EC）中的作用，为潜在的治疗靶点提供理论依据。通过生物信息学分析确定 EC 组织和正常组织中的差异表达基因。采用定量逆转录聚合酶链反应（qRT-PCR）比较正常人和 EC 细胞中 miR-18a-5p 和 THBD mRNA 的表达。CCK-8 法比较不同处理组 EC 细胞的增殖能力。Transwell 法检测不同处理组 EC 细胞的迁移和侵袭能力。双荧光素酶报告实验验证 miR-18a-5p 与 THBD 的靶向关系。Western blot 法检测 THBD 蛋白表达水平。qRT-PCR 结果显示，miR-18a-5p 在 EC 细胞中显著上调，其靶基因 THBD 的表达显著下调。CCK-8 和 Transwell 实验表明，miR-18a-5p 可增强 EC 细胞的增殖、迁移和侵袭能力，而 THBD 则可减弱这些能力。双荧光素酶报告实验证实 miR-18a-5p 可负调控 THBD 表达。此外，挽救实验表明 THBD 过表达可抑制 miR-18a-5p 对 EC 细胞的致癌作用。综上，miR-18a-5p 可能通过靶向下调 THBD 表达促进 EC 细胞的增殖、迁移和侵袭，miR-18a-5p/THBD 调控通路可能是一个治疗靶点。本研究结果可为相关药物研发提供理论依据。
