PDF(Pubmed)
Abstract：
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.
摘要：
本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。最初，采用细胞计数试剂盒-8(CCK8)试验严格研究姜黄素对TNBC细胞增殖能力的影响。随后，流式细胞术用于仔细评估姜黄素对细胞凋亡和细胞周期调节的影响。采用Transwell测定法仔细评估姜黄素对TNBC细胞运动性的影响。进行了RNA测序，其次是基因本体论和京都百科全书的基因和基因组富集分析差异表达基因，旨在阐明姜黄素潜在的抗癌机制。为了彻底阐明多种蛋白质之间的相互作用，我们构建了一个蛋白质-蛋白质相互作用(PPI)网络。最后,几种关键蛋白质的表达水平，包括纤连蛋白,mTOR,β-连环蛋白，p-Akt,Akt,N-钙黏着蛋白,使用蛋白质印迹评估p-S6和S6。CCK8检测结果显示,姜黄素显著抑制Hs578T和MDA-MB-231细胞的增殖。流式细胞术结果显示,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实验结果显示姜黄素有效降低Hs578T和MDA-MB-231细胞的运动性。RNA测序数据的富集分析表明，姜黄素的作用机制与肿瘤通路等信号通路显著相关，病灶粘连,和PI3K-Akt信号通路。随后，我们构建了一个蛋白质-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,Westernblotting分析表明,姜黄素显著降低纤维连接蛋白等关键蛋白的表达水平,mTOR,β-连环蛋白，p-Akt,Akt,N-钙黏着蛋白,p-S6和S6.姜黄素通过调节多种信号通路在TNBC中显示出其治疗潜力。它可能通过下调mTOR和PI3K-Akt信号通路相关蛋白的表达来抑制上皮间质转化过程,从而抑制TNBC细胞的运动性。这些发现为将姜黄素视为治疗TNBC的潜在治疗策略提供了实验证据。
