Institute of Medical Cancer, School of Basic Medical Sciences, Southwest Medical University, Luzhou, Sichuan, China.
The Second Clinical Medicine School, Hainan Medical College, Haikou, Hainan, China.
Medicine (Baltimore). 2024 Feb 23;103(8):e37264. doi: 10.1097/MD.0000000000037264.
This study aimed to use bioinformatics approaches for predicting the anticancer mechanisms of curcumin on triple-negative breast cancer (TNBC) and to verify these predictions through in vitro experiments. Initially, the Cell Counting Kit-8 (CCK8) assay was employed to rigorously investigate the influence of curcumin on the proliferative capacity of TNBC cells. Subsequently, flow cytometry was employed to meticulously assess the impact of curcumin on cellular apoptosis and the cell cycle regulation. Transwell assays were employed to meticulously evaluate the effect of curcumin on the motility of TNBC cells. RNA sequencing was conducted, followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses of differentially expressed genes, aiming to elucidate the potential anticancer mechanisms underlying curcumin's effects. To thoroughly elucidate the interactions among multiple proteins, we constructed a protein-protein interaction (PPI) network. Finally, the expression levels of several key proteins, including fibronectin, mTOR, β-Catenin, p-Akt, Akt, N-Cadherin, p-S6, and S6, were assessed using the western blot. The CCK8 assay results showed that curcumin significantly inhibited the proliferation of Hs578T and MDA-MB-231 cells. Flow cytometry results showed that curcumin induced apoptosis in these cells and arrested the cell cycle at the G2/M phase. Additionally, Transwell assay results showed that curcumin effectively reduced the motility of Hs578T and MDA-MB-231 cells. Enrichment analysis of RNA sequencing data showed that the mechanism of action of curcumin was significantly associated with signaling pathways such as pathways in cancer, focal adhesion, and PI3K-Akt signaling pathways. Subsequently, we constructed a protein-protein interaction network to elucidate the interactions among multiple proteins. Finally, Western blotting analysis showed that curcumin significantly decreased the expression levels of key proteins including Fibronectin, mTOR, β-Catenin, p-Akt, Akt, N-Cadherin, p-S6, and S6. Curcumin exhibits its therapeutic potential in TNBC by modulating multiple signaling pathways. It may inhibit the epithelial-mesenchymal transition process by downregulating the expression of proteins involved in the mTOR and PI3K-Akt signaling pathways, thereby suppressing the motility of TNBC cells. These findings provide experimental evidence for considering curcumin as a potential therapeutic strategy in the treatment of TNBC.
这项研究旨在利用生物信息学方法预测姜黄素对三阴性乳腺癌（TNBC）的抗癌机制，并通过体外实验验证这些预测。首先，采用细胞计数试剂盒-8（CCK8）实验严格研究姜黄素对 TNBC 细胞增殖能力的影响。随后，采用流式细胞术仔细评估姜黄素对细胞凋亡和细胞周期调控的影响。采用 Transwell 实验仔细评估姜黄素对 TNBC 细胞迁移能力的影响。进行 RNA 测序，并进行基因本体论和京都基因与基因组百科全书富集分析差异表达基因，旨在阐明姜黄素作用的潜在抗癌机制。为了彻底阐明多种蛋白质之间的相互作用，我们构建了蛋白质-蛋白质相互作用（PPI）网络。最后，使用 Western blot 评估几种关键蛋白，包括纤连蛋白、mTOR、β-连环蛋白、p-Akt、Akt、N-钙黏蛋白、p-S6 和 S6 的表达水平。CCK8 实验结果表明姜黄素显著抑制 Hs578T 和 MDA-MB-231 细胞的增殖。流式细胞术结果表明姜黄素诱导这些细胞凋亡并将细胞周期阻滞在 G2/M 期。此外，Transwell 实验结果表明姜黄素有效降低 Hs578T 和 MDA-MB-231 细胞的迁移能力。RNA 测序数据的富集分析表明，姜黄素的作用机制与癌症途径、黏附斑和 PI3K-Akt 信号通路等信号通路显著相关。随后，我们构建了蛋白质-蛋白质相互作用网络，以阐明多种蛋白质之间的相互作用。最后，Western blot 分析表明，姜黄素显著降低了纤连蛋白、mTOR、β-连环蛋白、p-Akt、Akt、N-钙黏蛋白、p-S6 和 S6 等关键蛋白的表达水平。姜黄素通过调节多条信号通路在 TNBC 中发挥治疗潜力。它可能通过下调参与 mTOR 和 PI3K-Akt 信号通路的蛋白表达来抑制上皮-间充质转化过程，从而抑制 TNBC 细胞的迁移。这些发现为将姜黄素作为治疗 TNBC 的潜在治疗策略提供了实验依据。
