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慢病毒介导的 microRNA-141 过表达促进人食管鳞癌细胞增殖并抑制细胞凋亡。

Lentiviral-Mediated Overexpression of MicroRNA-141 Promotes Cell Proliferation and Inhibits Apoptosis in Human Esophageal Squamous Cell Carcinoma.

机构信息

Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, Xi'an 710062, Shaanxi, China.

Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan, China.

出版信息

Recent Pat Anticancer Drug Discov. 2019;14(2):170-176. doi: 10.2174/1574892814666181231142136.

Abstract

BACKGROUND

Esophageal Carcinoma (EC) is the eighth most common cancer worldwide. Numerous studies have highlighted a vital role of microRNAs (miRNAs) in the development of EC. However, the mechanism of microRNA (miRNA)-141 in Esophageal Squamous Cell Carcinoma (ESCC) remains unknown.

OBJECTIVE

In this study, we explored the effects of miRNA-141 on EC cell proliferation, apoptosis, xenograft tumour growth and their possible mechanisms.

METHODS

A lentivirus-vector-expressing miRNA-141 was constructed, and a TE-1 cell line of ESCC with a stable expression of miRNA-141 was transfected and screened. The miRNA-141 expression level was detected using qRT-PCR. Effects of miRNA-141 overexpression on cell proliferation and apoptosis were detected using MTT and flow cytometry, respectively. Using a dual-luciferase reporter assay, a direct interaction between miRNA-141 and the 3'-Untranslated Region (UTR) of YAP1 and SOX17 was confirmed. Tumour xenograft experiment in nude mice was used to detect the tumour growth, and the effects of miRNA-141 overexpression on YAP1 and SOX17 were analysed using Western blot.

RESULTS

We found that miRNA-141 was highly expressed in TE-1 cells, and miRNA-141 overexpression promoted cell proliferation and inhibited apoptosis. Moreover, the miRNA-141 group showed significantly increased tumour growth ability, luciferase activities and expression levels of YAP1 and SOX17 in the miRNA-141group were significantly down-regulated.

CONCLUSION

miRNA-141 promotes cell proliferation and inhibits apoptosis in ESCC by downregulating the expression level of YAP1 and SOX17, indicating that miRNA-141 may be a potential molecular target for the treatment of ESCC.

摘要

背景

食管癌(EC)是全球第八大常见癌症。许多研究强调了 microRNAs(miRNAs)在 EC 发展中的重要作用。然而,microRNA(miRNA)-141 在食管鳞状细胞癌(ESCC)中的作用机制尚不清楚。

目的

本研究旨在探讨 miRNA-141 对 EC 细胞增殖、凋亡、异种移植肿瘤生长的影响及其可能的机制。

方法

构建表达 miRNA-141 的慢病毒载体,转染并筛选 ESCC 的 TE-1 细胞系,稳定表达 miRNA-141。采用 qRT-PCR 检测 miRNA-141 的表达水平。采用 MTT 和流式细胞术分别检测 miRNA-141 过表达对细胞增殖和凋亡的影响。采用双荧光素酶报告基因实验证实 miRNA-141 与 YAP1 和 SOX17 的 3'非翻译区(UTR)直接相互作用。裸鼠肿瘤异种移植实验检测肿瘤生长,采用 Western blot 分析 miRNA-141 过表达对 YAP1 和 SOX17 的影响。

结果

我们发现 miRNA-141 在 TE-1 细胞中高表达,miRNA-141 过表达促进细胞增殖,抑制细胞凋亡。此外,miRNA-141 组肿瘤生长能力显著增强,miRNA-141 组的荧光素酶活性和 YAP1、SOX17 的表达水平明显下调。

结论

miRNA-141 通过下调 YAP1 和 SOX17 的表达水平促进 ESCC 细胞增殖,抑制细胞凋亡,提示 miRNA-141 可能是 ESCC 治疗的潜在分子靶点。

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