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miRNA-144-5p 通过下调 CDCA3 调节肺腺癌细胞的增殖和凋亡。

MiRNA-144-5p down-modulates CDCA3 to regulate proliferation and apoptosis of lung adenocarcinoma cells.

机构信息

Thoracic Surgery, Zhejiang Hospital, Hangzhou, Zhejiang Province 310000, China.

Thoracic Surgery, Zhejiang Hospital, Hangzhou, Zhejiang Province 310000, China.

出版信息

Mutat Res. 2022 Jul-Dec;825:111798. doi: 10.1016/j.mrfmmm.2022.111798. Epub 2022 Sep 1.

Abstract

BACKGROUND

Lung adenocarcinoma (LUAD) belongs to non-small cell lung cancer. In addition to surgical resection, chemotherapy and radiotherapy cause great side effects and low 5-year survival rates. MiRNAs are closely related to cancer development. This study aimed to analyze the molecular mechanism of miRNA-144-5p targeting CDCA3 to inhibit LUAD proliferation.

METHODS

MiRNA and mRNA data were downloaded from TCGA-LUAD dataset for differential expression analysis. TargetScan and miRTarBase databases were adopted to predict the target genes of miRNA, and the signaling pathways involved were analyzed by gene set enrichment analysis. The functions of LUAD cells were analyzed by CCK-8, colony formation assay, stem cell spheroidization assay, and flow cytometry. The expression levels of CDCA3, p53, and cell cycle-associated proteins were evaluated by Western blot.

RESULTS

The expression of miRNA-144-5p was significantly down-regulated in LUAD, but overexpression of it repressed proliferation and spheroidization, and promoted apoptosis of LUAD cells. By bioinformatics prediction and dual-luciferase reporter assay, miRNA-144-5p was validated to target CDCA3, thereby regulating proliferation of LUAD cells. Besides, the results of cell experiments showed that miRNA-144-5p targeting CDCA3 affected cell proliferation and apoptosis in LUAD by regulating cell cycles, and miRNA-144-5p/CDCA3 mediated the p53 signaling pathway to affect the growth of LUAD cells.

SIGNIFICANCE

Through the study of the pathogenesis of miRNA-144-5p regulating LUAD, we can better understand the molecular mechanism underlying LUAD development.

摘要

背景

肺腺癌(LUAD)属于非小细胞肺癌。除了手术切除外,化疗和放疗会产生很大的副作用,且患者的 5 年生存率较低。miRNA 与癌症的发生发展密切相关。本研究旨在分析 miRNA-144-5p 靶向 CDCA3 抑制 LUAD 增殖的分子机制。

方法

从 TCGA-LUAD 数据集下载 miRNA 和 mRNA 数据进行差异表达分析。采用 TargetScan 和 miRTarBase 数据库预测 miRNA 的靶基因,并通过基因集富集分析分析其涉及的信号通路。通过 CCK-8、集落形成实验、干细胞球体形成实验和流式细胞术分析 LUAD 细胞的功能。通过 Western blot 评估 CDCA3、p53 和细胞周期相关蛋白的表达水平。

结果

miRNA-144-5p 在 LUAD 中的表达明显下调,但过表达可抑制其增殖和球体形成,并促进 LUAD 细胞凋亡。通过生物信息学预测和双荧光素酶报告基因实验验证了 miRNA-144-5p 靶向 CDCA3,从而调节 LUAD 细胞的增殖。此外,细胞实验结果表明,miRNA-144-5p 通过调控细胞周期影响 LUAD 细胞的增殖和凋亡,通过调节 p53 信号通路影响 LUAD 细胞的生长。

结论

通过研究 miRNA-144-5p 调控 LUAD 的发病机制,我们可以更好地了解 LUAD 发生发展的分子机制。

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