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miR-20a 通过 MTOR 信号通路抑制乳腺癌细胞增殖并促进其凋亡。

MiR-20a suppresses proliferation and facilitates apoptosis of breast cancer cells via the MTOR signaling pathway.

机构信息

Department of Ultrasound, The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Zhejiang, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Nov;24(22):11650-11657. doi: 10.26355/eurrev_202011_23809.

Abstract

OBJECTIVE

The paper aimed to explore the role of micro ribonucleic acid (miR)-20a in regulating the proliferation and apoptosis of breast cancer cells.

MATERIALS AND METHODS

The expression of miR-20a in breast cancer cells was analyzed via quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. Cell Counting Kit-8 (CCK-8) assay, colony formation assay, and flow cytometry were employed to analyze the proliferation and apoptosis of cells. Thereafter, the target proteins of miR-20a were predicted using TargetScan, a website for miRNA target gene prediction, and the interaction between miR-20a and the target genes was detected through the Luciferase reporter gene assay, qRT-PCR assay, and Western blotting. Finally, the miR-20a inhibitor and target gene expression plasmids were co-transfected for rescue experiment to study whether the target genes participate in the inhibitory effect of miR-20a on the proliferation of breast cancer cells.

RESULTS

It was found that the expression of miR-20a was upregulated in breast cancer cell lines. Silencing miR-20a expression inhibited the proliferation and promoted the apoptosis of breast cancer cell. Besides, it was demonstrated that late endosomal/lysosomal adaptor, mitogen-activated protein kinase (MAPK), and mammalian target of rapamycin (mTOR) activator 3 (LAMTOR3) were a direct target of miR-20a. The knockdown of LAMTOR3 expression repressed the influence of miR-20a on the proliferation of breast cancer cells.

CONCLUSIONS

MiR-20a targets LAMTOR3 gene to regulate the mTOR signaling pathway, thereby suppressing the proliferation and facilitating the apoptosis of breast cancer cells. It suggests that miR-20a exerts a carcinogenic effect in breast cancer, which may be a potential target for the treatment of breast cancer.

摘要

目的

本文旨在探讨微小 RNA(miR)-20a 在调节乳腺癌细胞增殖和凋亡中的作用。

材料和方法

采用实时定量聚合酶链反应(qRT-PCR)检测 miR-20a 在乳腺癌细胞中的表达。采用细胞计数试剂盒-8(CCK-8)检测、集落形成实验和流式细胞术分析细胞的增殖和凋亡。此后,利用 TargetScan 网站预测 miR-20a 的靶基因,通过荧光素酶报告基因检测、qRT-PCR 检测和 Western blot 检测 miR-20a 与靶基因的相互作用。最后,通过共转染 miR-20a 抑制剂和靶基因表达质粒进行挽救实验,研究靶基因是否参与 miR-20a 对乳腺癌细胞增殖的抑制作用。

结果

发现 miR-20a 在乳腺癌细胞系中表达上调。沉默 miR-20a 表达抑制乳腺癌细胞的增殖并促进其凋亡。此外,证明晚期内体/溶酶体衔接蛋白、丝裂原活化蛋白激酶(MAPK)和雷帕霉素靶蛋白(mTOR)激活物 3(LAMTOR3)是 miR-20a 的直接靶基因。LAMTOR3 表达的敲低抑制了 miR-20a 对乳腺癌细胞增殖的影响。

结论

miR-20a 靶向 LAMTOR3 基因调控 mTOR 信号通路,从而抑制乳腺癌细胞的增殖并促进其凋亡。这表明 miR-20a 在乳腺癌中发挥致癌作用,可能成为治疗乳腺癌的潜在靶点。

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