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微小 RNA miR-124-3p 通过 ARRDC1(含有抑制素结构域蛋白 1)抑制肝癌的增殖和上皮-间充质转化。

MicroRNA miR-124-3p suppresses proliferation and epithelial-mesenchymal transition of hepatocellular carcinoma via ARRDC1 (arrestin domain containing 1).

机构信息

Department of Laboratory Medicine, Yantai Mountain Hospital, Yantai, Shandong, China.

Health Management Center, Qishan Hospital, Yantai, Shandong, China.

出版信息

Bioengineered. 2022 Apr;13(4):8255-8265. doi: 10.1080/21655979.2022.2051686.

Abstract

Hepatocellular carcinoma (HCC) is responsible for high morbidity and mortality worldwide. Increasing evidence suggests that microRNAs intensively participate in HCC development and progression. In the current study, we aimed to explore the impact of miR-124-3p in the proliferation and epithelial-mesenchymal transition (EMT) of HCC. The RT-qPCR assay was employed to determine miR-124-3p expression in human HCC specimens and cell lines. Luciferase assay was used to validate the miR-124-3p target gene. Western Blot and RT-qPCR were performed to study the effects of miR-124-3p modulation on ARRDC1 (Arrestin Domain Containing 1) mRNA and protein expressions. MTT assay, wound healing assay, EdU assay, and Transwell assay were utilized to verify the impact of miR-144-3p modulation on HCC proliferation and EMT via ARRDC1. We found that MiR-124-3p expression downregulates in HCC. Overexpression of miR-124-3p reduced the HCC cell proliferation and EMT. Meanwhile, we determined that the expression of ARRDC1 is increased in HCC, and miR-124-3p directly binds the 3'UTR of ARRDC1 and inhibits its expression at mRNA and protein level, suggesting that miR-124-3p was capable of negatively modulating ARRDC1. Besides, cotransfection of ARRDC1-overexpression plasmid and miR-124-3p mimics increased the cell proliferation and EMT as compared to miR-124-3p mimics. Our study concluded that miR-124-3p directly binds the 3'UTR of ARRDC1 and exerts anti-tumorous effects by inhibiting the HCC proliferation and EMT. Therefore, miR-124-3p/ARRDC1 axis may serve as a novel therapeutic target to inhibit HCC growth and metastasis.

摘要

肝细胞癌 (HCC) 是全球发病率和死亡率较高的疾病。越来越多的证据表明,miRNA 广泛参与 HCC 的发生和发展。在本研究中,我们旨在探讨 miR-124-3p 在 HCC 增殖和上皮间质转化 (EMT) 中的作用。采用 RT-qPCR 法检测人 HCC 标本和细胞系中 miR-124-3p 的表达。荧光素酶报告基因实验验证 miR-124-3p 的靶基因。Western blot 和 RT-qPCR 检测 miR-124-3p 调控 ARRDC1 (Arrestin Domain Containing 1) mRNA 和蛋白表达的影响。MTT 试验、划痕愈合试验、EdU 试验和 Transwell 试验验证 miR-124-3p 通过 ARRDC1 对 HCC 增殖和 EMT 的影响。结果发现,miR-124-3p 在 HCC 中表达下调。miR-124-3p 的过表达降低了 HCC 细胞的增殖和 EMT。同时,我们发现 ARRDC1 在 HCC 中的表达增加,miR-124-3p 可直接结合 ARRDC1 的 3'UTR,抑制其在 mRNA 和蛋白水平的表达,表明 miR-124-3p 能够负调控 ARRDC1。此外,与 miR-124-3p 模拟物转染相比,共转染 ARRDC1 过表达质粒和 miR-124-3p 模拟物增加了细胞增殖和 EMT。综上所述,miR-124-3p 可直接结合 ARRDC1 的 3'UTR,通过抑制 HCC 增殖和 EMT 发挥抑癌作用。因此,miR-124-3p/ARRDC1 轴可能成为抑制 HCC 生长和转移的新治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c148/9161870/e0582f73acdb/KBIE_A_2051686_UF0001_OC.jpg

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