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miRNA-96-5p 通过靶向 FOXO3 影响乳腺癌的进展。

MiRNA-96-5p impacts the progression of breast cancer through targeting FOXO3.

机构信息

Department of Breast Surgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.

Department of Medical Oncology, Xuzhou Central Hospital, Xuzhou, China.

出版信息

Thorac Cancer. 2020 Apr;11(4):956-963. doi: 10.1111/1759-7714.13348. Epub 2020 Feb 26.

Abstract

BACKGROUND

Breast cancer is the most common malignant tumor in women worldwide, with a high mortality rate. MicroRNAs are small non-coding RNAs that negatively regulate the expression of target genes by interacting with the target gene 3'-UTR, and participate in cell differentiation, proliferation, apoptosis and metabolism. The function of miRNA-96-5p in the progression of breast cancer has not been reported.

METHODS

We used the StarBase database to investigate the expression of miRNA-96-5p in breast cancer and adjacent normal tissues. FOXO3 3'-UTR construct and luciferase reporter assays was performed for the target gene. Expression levels of miRNAs including its target were analyzed by qRT-PCR and western blot. Cell proliferation was detected by CCK8 and colony formation, EdU assay.

RESULTS

Luciferase reporter assays showed miRNA-96-5p directly targeted FOXO3. Abrogation of miRNA-96-5p by transfection with its inhibitors in breast cancer cells significantly suppressed miRNA-96-5p expression and breast cancer cells proliferation. Western blot revealed that overexpression of miRNA-96-5p substantially reduced FOXO3 protein expression. We used the GEPIA, UALCAN and KM-plotter databases to investigate the expression of FOXO3 in human breast cancer and adjacent normal tissues, and its correlation with survival. In addition, we found that FOXO3 spoiled miR-96-5p induced breast cancer cell proliferation block effecting.

CONCLUSIONS

miRNA-96-5p may exert a tumor promotion role through negatively regulating tumor suppressor gene FOXO3 and promoting cell proliferation.

摘要

背景

乳腺癌是全球女性中最常见的恶性肿瘤,死亡率较高。miRNA 是一种小的非编码 RNA,通过与靶基因 3'-UTR 相互作用负调控靶基因的表达,参与细胞分化、增殖、凋亡和代谢。miRNA-96-5p 在乳腺癌进展中的作用尚未报道。

方法

我们使用 StarBase 数据库研究 miRNA-96-5p 在乳腺癌及相邻正常组织中的表达。构建 FOXO3 3'-UTR 载体和荧光素酶报告基因检测其靶基因。通过 qRT-PCR 和 Western blot 分析 miRNA 及其靶基因的表达水平。通过 CCK8 法和集落形成实验、EdU 实验检测细胞增殖。

结果

荧光素酶报告基因实验显示 miRNA-96-5p 可直接靶向 FOXO3。用其抑制剂转染乳腺癌细胞可显著抑制 miRNA-96-5p 的表达并抑制乳腺癌细胞的增殖。Western blot 显示过表达 miRNA-96-5p 可显著降低 FOXO3 蛋白的表达。我们使用 GEPIA、UALCAN 和 KM-plotter 数据库研究了 FOXO3 在人乳腺癌及相邻正常组织中的表达及其与生存的关系。此外,我们发现 FOXO3 可削弱 miR-96-5p 诱导的乳腺癌细胞增殖阻滞作用。

结论

miRNA-96-5p 可能通过负调控抑癌基因 FOXO3 促进细胞增殖,发挥促肿瘤作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a7e5/7113053/eb4253bb69c4/TCA-11-956-g001.jpg

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