Ankara University, Biotechnology Institute, Ankara, Turkey.
Hum Exp Toxicol. 2020 Nov;39(11):1497-1506. doi: 10.1177/0960327120930257. Epub 2020 Jun 8.
This study aims to determine whether usnic acid (UA) could induce the expression of apoptosis-related genes in apoptosis pathway. The current study has enabled us to better understand the target of UA in the treatment of breast cancer. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Based on the previous study and the results of this study, UA had the most antiproliferative effect on SK-BR-3 breast cancer cell line. We examined differential expression of 88 apoptosis-related genes by quantitative real-time polymerase chain reaction using the apoptosis primary library panel in SK-BR-3 breast cancer cell. We observed a difference in the significant differential expression of 74 apoptosis-related genes in breast cancer after SK-BR-3 cells applied to UA (7.21 µM) for 48 h. The expression level of 56 of these 74 differentiated apoptosis-related genes increased ( < 0.05), but the expression level of the other 18 related genes decreased ( < 0.05). In order to evaluate the mechanism of apoptosis of UA, Western blot analysis was performed with Bcl-2, Bax, Caspase-3, and Caspase-9 antibodies. According to the Western blot analysis, we obtained similar results with gene-expression data. These results suggest that UA showed a cytotoxic effect in SK-BR-3 cells through activation of the mitochondrial apoptotic pathway. The obtained results from gene expression revealed that the effect of UA on apoptosis pathway is critical for clinical research.
本研究旨在确定乌头酸(UA)是否能诱导凋亡途径中凋亡相关基因的表达。目前的研究使我们能够更好地了解 UA 在治疗乳腺癌中的作用靶点。细胞活力通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法测定。基于先前的研究和本研究的结果,UA 对 SK-BR-3 乳腺癌细胞系的增殖抑制作用最明显。我们通过定量实时聚合酶链反应使用凋亡初级文库面板在 SK-BR-3 乳腺癌细胞中检查了 88 个凋亡相关基因的差异表达。我们观察到 SK-BR-3 细胞应用 UA(7.21 µM)48 小时后,乳腺癌中 74 个凋亡相关基因的差异表达有显著差异。这些差异表达的 74 个凋亡相关基因中的 56 个基因的表达水平升高(<0.05),但另外 18 个相关基因的表达水平降低(<0.05)。为了评估 UA 诱导凋亡的机制,我们用 Bcl-2、Bax、Caspase-3 和 Caspase-9 抗体进行了 Western blot 分析。根据 Western blot 分析,我们得到了与基因表达数据相似的结果。这些结果表明,UA 通过激活线粒体凋亡途径在 SK-BR-3 细胞中表现出细胞毒性作用。基因表达获得的结果表明,UA 对凋亡途径的作用对临床研究至关重要。