三阴性乳腺癌（TNBC）是一种乳腺癌亚型，其具有预后差和治疗选择有限的特点。橄榄苦苷和橄榄环醇是在特级初榨橄榄油中发现的生物活性化合物，它们已被证明具有抗癌潜力。在本研究中，我们检测了这两种天然化合物对MDA-MB-231和MDA-MB-468 TNBC细胞系的抑制作用。将人体TNBC MDA-MB-231和MDA-MB-468细胞系分别用橄榄苦苷或橄榄环醇进行不同浓度的处理48小时。使用硫酸罗丹明B（sulforhodamine B）测定法确定达到IC50的最佳浓度后，在处理和未处理的细胞中分别提取12、24和48小时后的总RNA。进行聚A RNA筛选，然后进行文库构建和RNA测序。进行差异基因表达（DEG）分析，以确定处理和未处理细胞之间的DEG。使用KEGG和GO数据库进行通路分析。橄榄苦苷和橄榄环醇显著抑制了TNBC细胞的增殖，其中橄榄环醇的效果略大于橄榄苦苷。此外，多时间序列RNA测序显示，在处理这些化合物后，TNBC细胞的表达谱发生了显著改变，有些基因在所有时间点上都一直受到了时间动力学和基因组的影响。通路分析揭示了与TNBC相关的几个重要通路，包括细胞死亡、凋亡过程、程序性细胞死亡、应激反应、有丝分裂细胞周期过程、细胞分裂和癌症进程。我们的研究结果表明，橄榄苦苷和橄榄环醇对TNBC具有潜在的治疗好处，并可进一步作为替代治疗选择进行深入研究。© 2023 作者。

Triple-negative breast cancer (TNBC) is a subtype of breast cancer characterized by poor prognosis and limited treatment options. Oleuropein and oleocanthal are bioactive chemicals found in extra-virgin olive oil; they have been shown to have anti-cancer potential. In this study, we examined the inhibitory effects of these two natural compounds, on MDA-MB-231 and MDA-MB-468 TNBC cell lines. The human TNBC MDA-MB-231 and MDA-MB-468 cell lines were treated with oleuropein or oleocanthal at ranging concentrations for 48 h. After determining the optimum concentration to reach IC50, using the sulforhodamine B assay, total RNA was extracted after 12, 24, and 48 h from treated and untreated cells. Poly(A)-RNA selection was conducted, followed by library construction and RNA sequencing. Differential gene expression (DEG) analysis was performed to identify DEGs between treated and untreated cells. Pathway analysis was carried out using the KEGG and GO databases. Oleuropein and oleocanthal considerably reduced the proliferation of TNBC cells, with oleocanthal having a slightly stronger effect than oleuropein. Furthermore, multi-time series RNA sequencing showed that the expression profile of TNBC cells was significantly altered after treatment with these compounds, with temporal dynamics and groups of genes consistently affected at all time points. Pathway analysis revealed several significant pathways associated with TNBC, including cell death, apoptotic process, programmed cell death, response to stress, mitotic cell cycle process, cell division, and cancer progression. Our findings suggest that oleuropein and oleocanthal have potential therapeutic benefits for TNBC and can be further investigated as alternative treatment options.© 2023. The Author(s).