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长链非编码 RNA FERRE 的上调通过调节 miR-19a-5p/EZH2 轴促进乳腺癌的生长和侵袭。

Upregulation of long noncoding RNA FERRE promoted growth and invasion of breast cancer through modulating miR-19a-5p/EZH2 axis.

机构信息

Institute for Cancer Medicine, School of Basic Medical Sciences, Southwest Medical University, Luzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Nov;24(21):11154-11164. doi: 10.26355/eurrev_202011_23603.

Abstract

OBJECTIVE

It has been demonstrated that long non-coding RNA (LncRNA) plays an important regulatory role in a series of diseases. The purpose of this study is to investigate the expression of long non-coding RNA (LncRNA) FERRE and its facilitating effects on proliferation and invasion of breast cancer by regulating oncogene EZH2 through sponging with miR-19a-5p.

PATIENTS AND METHODS

qRT-PCR was performed to detect the expressions of FERRE and EZH2 in human breast cancer tissues and cells. CCK-8 assay was performed to evaluate the MCF-7 cells proliferation and transwell assay was performed to evaluate the MCF-7 cells migration. Correlation analysis between FERRE and miR-19a-5p was detected by statistical analysis. Bioinformatics prediction was made to detect the binding site of FERRE and miR-19a-5p and Luciferase activity was conducted to investigate the interaction between EZH2 and miR-19a-5p. Furthermore, we cloned the mice EZH2 3'-UTR into the Luciferase reporter vector and constructed miR-19a-5p binding mutants to validate the inhibited modulation of miR-19a-5p to the EZH2 expression.

RESULTS

Results showed that expression of FERRE and EZH2 were upregulated in human breast cancer tissues and cells. qRT-PCR and CCK-8 assay showed that FERRE expression is associated with the proliferation of breast cancer cells, upregulated FERRE contributed to cell proliferation of MCF-7. Transwell assay showed that FERRE was associated with the migration ability of tumor cells, increased expression of FERRE promoted the migration and invasion of breast cancer cells. The bioinformatics prediction and Luciferase assay demonstrated that by sponging with miR-19a-5p, FERRE can serve as a molecular sponge to further regulate the expression of EZH2.

CONCLUSIONS

We found that lncRNA-FERRE was upregulated in human breast cancer patients, which could accelerate tumor proliferation, migration and invasion as a molecular sponge by modulating the inhibitory effect of miR-19a-5p on oncogene EZH2.

摘要

目的

长链非编码 RNA(LncRNA)在一系列疾病中发挥重要的调控作用已被证实。本研究旨在通过海绵吸附 miR-19a-5p 调控癌基因 EZH2,探讨长链非编码 RNA(LncRNA)FERRE 的表达及其促进乳腺癌增殖和侵袭的作用。

患者和方法

采用 qRT-PCR 检测人乳腺癌组织和细胞中 FERRE 和 EZH2 的表达。CCK-8 法检测 MCF-7 细胞增殖,Transwell 法检测 MCF-7 细胞迁移。统计分析检测 FERRE 与 miR-19a-5p 的相关性。生物信息学预测 FERRE 与 miR-19a-5p 的结合位点,采用荧光素酶活性实验检测 EZH2 与 miR-19a-5p 的相互作用。进一步将小鼠 EZH2 3'-UTR 克隆到荧光素酶报告载体中,构建 miR-19a-5p 结合突变体,验证 miR-19a-5p 对 EZH2 表达的抑制调节作用。

结果

结果表明,FERRE 和 EZH2 在人乳腺癌组织和细胞中表达上调。qRT-PCR 和 CCK-8 实验显示,FERRE 的表达与乳腺癌细胞的增殖有关,上调 FERRE 促进 MCF-7 细胞的增殖。Transwell 实验显示,FERRE 与肿瘤细胞的迁移能力有关,增加 FERRE 的表达促进乳腺癌细胞的迁移和侵袭。生物信息学预测和荧光素酶实验表明,通过海绵吸附 miR-19a-5p,FERRE 可以作为一种分子海绵,进一步调节癌基因 EZH2 的表达。

结论

我们发现,lncRNA-FERRE 在人乳腺癌患者中上调,作为一种分子海绵,通过调节 miR-19a-5p 对癌基因 EZH2 的抑制作用,加速肿瘤的增殖、迁移和侵袭。

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