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miR-148a-3p 通过靶向 DNMT1 抑制食管癌的增殖和侵袭。

miR-148a-3p Suppresses the Proliferation and Invasion of Esophageal Cancer by Targeting DNMT1.

作者信息

Wang Yuping, Hu Yuna, Guo Junhui, Wang Lingling

机构信息

First Oncology Ward, Henan Province Hospital of TCM, Zhengzhou, P.R. China.

出版信息

Genet Test Mol Biomarkers. 2019 Feb;23(2):98-104. doi: 10.1089/gtmb.2018.0285.

Abstract

AIM

To identify whether miR-148a-3p interacts with DNA (cytosine-5)-methyltransferase 1 (DNMT1) in esophageal cancer.

METHODS

A luciferase assay and immunoblotting were performed to detect the relationship between miR-148a-3p and DNMT1. The MTT method, Annexin V/propidium iodide staining, and Transwell assays were adopted to assess the biological behaviors in EC109 cells. The association between the expression level of miR-148a-3p, clinical features, and prognosis were evaluated by chi-square test and univariate survival analysis.

RESULTS

In this study, DNMT1 was identified as a direct target of miR-148a-3p by luciferase assay and Western blot. Real-time quantitative PCR analyses showed that the relative expression levels of miR-148a-3p and DNMT1 were reduced in esophageal cancer samples compared with adjacent tissues; and a negative relationship between both was indicated. Upon overexpression of miR-148a-3p in esophageal cancer cells, proliferation and invasion were significantly suppressed, and apoptosis was promoted. A higher level of miR-148a-3p was correlated with better patient outcomes.

CONCLUSIONS

Our study indicated that miR-148a-3p, by targeting DNMT1, likely regulates cell proliferation and invasion in esophageal cancer. miR-148a-3p might also be used prognostically in esophageal cancer and serve as a therapeutic target in the future.

摘要

目的

确定在食管癌中miR-148a-3p是否与DNA(胞嘧啶-5)-甲基转移酶1(DNMT1)相互作用。

方法

进行荧光素酶报告基因检测和免疫印迹以检测miR-148a-3p与DNMT1之间的关系。采用MTT法、膜联蛋白V/碘化丙啶染色和Transwell实验评估EC109细胞的生物学行为。通过卡方检验和单因素生存分析评估miR-148a-3p表达水平与临床特征及预后之间的关联。

结果

在本研究中,通过荧光素酶报告基因检测和蛋白质免疫印迹确定DNMT1是miR-148a-3p的直接靶点。实时定量PCR分析显示,与癌旁组织相比,食管癌样本中miR-148a-3p和DNMT1的相对表达水平降低;且二者呈负相关。在食管癌细胞中过表达miR-148a-3p后,细胞增殖和侵袭受到显著抑制,细胞凋亡增加。miR-148a-3p水平较高与患者较好的预后相关。

结论

我们的研究表明,miR-148a-3p通过靶向DNMT1可能调控食管癌细胞的增殖和侵袭。miR-148a-3p也可能在食管癌预后评估中发挥作用,并有望成为未来的治疗靶点。

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