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长链非编码 RNA ZFAS1 的下调通过调节 miR-302-3p 对细胞周期蛋白 D1 的表达抑制甲状腺癌的标志性特征。

Downregulation of lncRNA ZFAS1 inhibits the hallmarks of thyroid carcinoma via the regulation of miR‑302‑3p on cyclin D1.

机构信息

Department of Pathology, The First Hospital of Qiqihar, Affiliated Qiqihar Hospital, Southern Medical University, Qiqihar, Heilongjiang 161000, P.R. China.

Department of Thyroid Surgery, The First Hospital of Qiqihar, Affiliated Qiqihar Hospital, Southern Medical University, Qiqihar, Heilongjiang 161000, P.R. China.

出版信息

Mol Med Rep. 2021 Jan;23(1). doi: 10.3892/mmr.2020.11640. Epub 2020 Nov 12.

Abstract

At present, treatment options for thyroid carcinoma remain limited. The present study aimed to investigate the role of ZFAS1 in various major hallmarks of cancer and the underlying mechanisms in thyroid carcinoma cells. The interactions between long non‑coding RNAs (lncRNAs), microRNAs (miRs) and target genes were predicted by bioinformatics and confirmed by performing dual‑luciferase assays. The mRNA and protein expressions were determined by reverse transcription‑quantitative PCR and western blotting. Cell invasion, migration, and viability were evaluated via Transwell, wound‑healing and Cell Counting Kit‑8 assays, respectively. The results demonstrated that lncRNA ZFAS1 expression was upregulated in thyroid carcinoma tissues, TT and SW579 cells, and was associated with the proliferation of these two cell lines. Notably, downregulation ZFAS1 reduced migration and invasion, and reversed the promotive effects of miR‑302a‑3p inhibitor on the proliferation, migration and invasion of TT and SW579 cells. Moreover, cyclin D1 (CCND1) is targeted by miR‑302a‑3p, and was regulated by ZFAS1. In addition, the downregulation of ZFAS1 not only reversed the promotive effects of miR‑302a‑3p inhibitor on CCND1 expression and the epithelial‑mesenchymal transition (EMT) of TT and SW579 cells, but also targeted and increased the expression of miR‑302a‑3p, and further reduced the expression of CCND1, resulting in suppression of the proliferation, migration, invasion and EMT of thyroid carcinoma cells.

摘要

目前,甲状腺癌的治疗选择仍然有限。本研究旨在探讨 ZFAS1 在甲状腺癌细胞中各种主要癌症特征中的作用及其潜在机制。通过生物信息学预测长链非编码 RNA(lncRNA)、微小 RNA(miR)与靶基因之间的相互作用,并通过双荧光素酶报告基因实验进行验证。采用反转录定量 PCR 和 Western blot 检测 mRNA 和蛋白表达。通过 Transwell 迁移实验、划痕愈合实验和细胞计数试剂盒-8(Cell Counting Kit-8,CCK-8)检测细胞侵袭、迁移和活力。结果表明,甲状腺癌组织和 TT、SW579 细胞中 lncRNA ZFAS1 的表达上调,与这两种细胞系的增殖相关。值得注意的是,下调 ZFAS1 可降低迁移和侵袭能力,并逆转 miR-302a-3p 抑制剂对 TT 和 SW579 细胞增殖、迁移和侵袭的促进作用。此外,miR-302a-3p 靶向调控细胞周期蛋白 D1(cyclin D1,CCND1),而 ZFAS1 可调节 CCND1 的表达。此外,下调 ZFAS1 不仅逆转了 miR-302a-3p 抑制剂对 TT 和 SW579 细胞中 CCND1 表达和上皮间质转化(epithelial-mesenchymal transition,EMT)的促进作用,还靶向并增加了 miR-302a-3p 的表达,进一步降低了 CCND1 的表达,从而抑制了甲状腺癌细胞的增殖、迁移、侵袭和 EMT。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/948c/7673324/df787f02e2e9/mmr-23-01-11640-g00.jpg

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