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长链非编码 RNA ZEB1-AS1 通过靶向 miR-299-3p/E2F1 轴促进肝癌细胞的增殖和转移。

Long non-coding RNA ZEB1-AS1 promotes proliferation and metastasis of hepatocellular carcinoma cells by targeting miR-299-3p/E2F1 axis.

机构信息

Department of Tenth Liver Disease, Qingdao Sixth People's Hospital, 9 Fushun Road, Qingdao 266033, Shandong Province, China.

Department of Medical Laboratory, Qingdao Sixth People's Hospital, 9 Fushun Road, Qingdao 266033, Shandong Province, China.

出版信息

J Biochem. 2021 Sep 22;170(1):41-50. doi: 10.1093/jb/mvab042.

Abstract

There is emerging evidence that dysregulation of long non-coding RNAs (lncRNAs) is associated with hepatocellular carcinoma (HCC). Zinc finger E-box binding homeobox 1 antisense 1 (ZEB1-AS1) functions as an oncogenic regulator in various malignancies. Nonetheless, the potential role of ZEB1-AS1 in HCC remains poorly elucidated. Herein, qRT-PCR was employed for examining ZEB1-AS1, miR-299-3p and E2F transcription factor 1 (E2F1) mRNA expressions in HCC cells and tissues. MTT assay was performed to evaluate cell proliferation. Transwell assay was utilized for evaluating cancer cell migration and invasion. Western blot was employed for measuring E2F1 protein expression. What's more, dual-luciferase reporter assay was utilized for verifying the targeting relationships between ZEB1-AS1 and miR-299-3p, as well as E2F1 and miR-299-3p. It was demonstrated that, in HCC tissues and cells, ZEB1-AS1 expression was markedly increased, and meanwhile, its high expression level is related to the unfavourable clinicopathologic indicators. ZEB1-AS1 overexpression facilitated HCC cell proliferation, migration and invasion, while its knockdown led to the opposite effects. In terms of mechanism, we discovered that ZEB1-AS1 could decoy miR-299-3p and upregulate E2F1 expression. This work reveals the functions and mechanism of ZEB1-AS1 in HCC tumourigenesis and progression, which provides novel biomarkers and therapeutic targets for HCC.

摘要

越来越多的证据表明,长非编码 RNA(lncRNA)的失调与肝细胞癌(HCC)有关。锌指 E 盒结合同源盒 1 反义 1(ZEB1-AS1)在多种恶性肿瘤中作为致癌调节剂发挥作用。然而,ZEB1-AS1 在 HCC 中的潜在作用仍未得到充分阐明。在此,我们通过 qRT-PCR 检测了 HCC 细胞和组织中的 ZEB1-AS1、miR-299-3p 和 E2F 转录因子 1(E2F1)mRNA 的表达。MTT 试验用于评估细胞增殖。Transwell 试验用于评估癌细胞的迁移和侵袭。Western blot 用于测量 E2F1 蛋白的表达。此外,我们还利用双荧光素酶报告基因试验验证了 ZEB1-AS1 与 miR-299-3p 之间,以及 E2F1 与 miR-299-3p 之间的靶向关系。结果表明,在 HCC 组织和细胞中,ZEB1-AS1 的表达显著增加,同时,其高表达水平与不良的临床病理指标有关。ZEB1-AS1 的过表达促进了 HCC 细胞的增殖、迁移和侵袭,而其敲低则导致相反的效果。在机制方面,我们发现 ZEB1-AS1 可以结合 miR-299-3p 并上调 E2F1 的表达。这项工作揭示了 ZEB1-AS1 在 HCC 肿瘤发生和进展中的功能和机制,为 HCC 提供了新的生物标志物和治疗靶点。

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