Zhang Fengfeng, Wang Binbin, Long Houlong, Yu Jingkui, Li Feng, Hou Haitao, Yang Qifeng
Clin Lab. 2016;62(6):1139-45. doi: 10.7754/clin.lab.2015.151111.
Recent findings have revealed that abnormal expression of microRNAs (miRNA, miR) contributes to the malignancies of various cancers. Here, we report a novel miRNA that regulates the expression of Beclin-1 in breast cancer cells.
The expression of miR-124-3p and Beclin-1 was identified in breast cancer tissues and breast cancer cell lines. To explore whether Beclin-1 was the target gene of miR-124-3p, luciferase reporter assay was applied. MIR-124-3p was overexpressed or inhibited with the corresponding mimics or inhibitors. The expression of autophagy-related proteins including Beclin-1 and LC3II were explored by western blot and quantitative real-time PCR.
We first demonstrated that miR-124-3p was decreased in breast cancer tissues and breast cancer cells lines. Furthermore, we validated that miR-124-3p could negatively regulate the expression of Beclin-1. Increased miR-124-3p significantly decreased the expression of Beclin-1 and LC3I. Further study showed that overexpres- sion of miR-124-3p could partially reverse 4-hydroxytamoxifen (4-OHT)-induced autophagy in breast cancer cells.
Decreased miR-124-3p expression prompted breast cancer cell progression mainly by enhancing the expression of autophagy related protein, Beclin-1.
最近的研究发现表明,微小RNA(miRNA,miR)的异常表达与多种癌症的恶性肿瘤发生有关。在此,我们报告一种新型的miRNA,其可调节乳腺癌细胞中Beclin-1的表达。
检测乳腺癌组织和乳腺癌细胞系中miR-124-3p和Beclin-1的表达。为探究Beclin-1是否为miR-124-3p的靶基因,采用荧光素酶报告基因检测法。用相应的模拟物或抑制剂过表达或抑制miR-124-3p。通过蛋白质免疫印迹法和定量实时聚合酶链反应检测包括Beclin-1和LC3II在内的自噬相关蛋白的表达。
我们首先证明miR-124-3p在乳腺癌组织和乳腺癌细胞系中表达降低。此外,我们证实miR-124-3p可负向调节Beclin-1的表达。miR-124-3p表达增加显著降低了Beclin-1和LC3I的表达。进一步研究表明,miR-124-3p过表达可部分逆转4-羟基他莫昔芬(4-OHT)诱导的乳腺癌细胞自噬。
miR-124-3p表达降低主要通过增强自噬相关蛋白Beclin-1的表达促进乳腺癌细胞进展。