Department of Ultrasound, The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Zhejiang, China.
Eur Rev Med Pharmacol Sci. 2020 Nov;24(22):11650-11657. doi: 10.26355/eurrev_202011_23809.
The paper aimed to explore the role of micro ribonucleic acid (miR)-20a in regulating the proliferation and apoptosis of breast cancer cells.
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.
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.
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.
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。
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。
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。
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。