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微小 RNA-488 通过直接靶向 ATF3 抑制舌鳞癌细胞侵袭和 EMT。

MicroRNA-488 inhibits tongue squamous carcinoma cell invasion and EMT by directly targeting ATF3.

机构信息

Oral and Maxillofacial Surgery, Cangzhou Central Hospital, No. 16 Xinhua West Road, Cangzhou, Hebei 061000 People's Republic of China.

出版信息

Cell Mol Biol Lett. 2018 Jun 18;23:28. doi: 10.1186/s11658-018-0094-0. eCollection 2018.

Abstract

BACKGROUND

It has been reported that the expression of activating transcription factor 3 (ATF3) is closely associated with both microRNA (miRNA) processing and the progress of many cancers. Our study aimed to explore the interaction between ATF3 and miR-488 in tongue squamous cell carcinoma (TSCC).

METHODS

Quantitative real-time PCR was performed to detect the levels of ATF3 and miR-488 in TSCC tissues and cell lines. Cell invasion and epithelial-mesenchymal transition (EMT) were assessed to determine the biological functions of miR-488 and ATF3 in TSCC cells. The mRNA and protein levels of ATF3 were measured using quantitative RT-PCR and western blotting. Luciferase assays were performed to validate ATF3 as an miR-488 target in TSCC cells.

RESULTS

We found that the level of miR-488 significantly decreased and the expression of ATF3 significantly increased in TSCC tissues and cell lines. A low level of miR-488 was closely associated with increased expression of ATF3 in TSCC tissues. Introducing miR-488 significantly inhibited the invasion and EMT of TSCC cells, and knockdown of miR-488 promoted both processes. The bioinformatics analysis predicted that ATF3 is a potential target gene of miR-488. The luciferase reporter assay showed that miR-488 could directly target ATF3. ATF3 silencing had similar effects to miR-488 overexpression on TSCC cells. Overexpression of ATF3 in TSCC cells partially reversed the inhibitory effects of the miR-488 mimic.

CONCLUSION

miR-488 inhibited cell invasion and EMT of TSCC cells by directly downregulating ATF3 expression.

摘要

背景

已有报道称,激活转录因子 3(ATF3)的表达与 microRNA(miRNA)加工和多种癌症的进展密切相关。本研究旨在探讨 ATF3 与舌鳞状细胞癌(TSCC)中 miR-488 的相互作用。

方法

采用实时定量 PCR 检测 TSCC 组织和细胞系中 ATF3 和 miR-488 的水平。通过细胞侵袭和上皮-间充质转化(EMT)评估来确定 miR-488 和 ATF3 在 TSCC 细胞中的生物学功能。采用定量 RT-PCR 和 Western blot 检测 ATF3 的 mRNA 和蛋白水平。通过荧光素酶报告基因实验验证 ATF3 是 TSCC 细胞中 miR-488 的靶基因。

结果

我们发现,miR-488 的水平在 TSCC 组织和细胞系中显著降低,而 ATF3 的表达显著升高。在 TSCC 组织中,miR-488 水平低与 ATF3 表达增加密切相关。引入 miR-488 可显著抑制 TSCC 细胞的侵袭和 EMT,而 miR-488 的敲低则促进了这两个过程。生物信息学分析预测 ATF3 是 miR-488 的潜在靶基因。荧光素酶报告基因实验表明,miR-488 可以直接靶向 ATF3。ATF3 沉默对 TSCC 细胞的作用与 miR-488 过表达相似。在 TSCC 细胞中过表达 ATF3 部分逆转了 miR-488 模拟物的抑制作用。

结论

miR-488 通过直接下调 ATF3 表达抑制 TSCC 细胞的侵袭和 EMT。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a159/6006839/a11c8112cd3f/11658_2018_94_Fig1_HTML.jpg

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