Kharazi Dania Mahmoud, Karam Louna, El Boustany Charbel, Ibrahim José-Noel
Department of Biological Sciences, School of Arts and Sciences, Lebanese American University (LAU), Beirut, Lebanon.
Department of Laboratory Science, Faculty of Public Health - Branch 2, Lebanese University, Fanar, Lebanon.
Front Cell Dev Biol. 2025 Mar 12;13:1535563. doi: 10.3389/fcell.2025.1535563. eCollection 2025.
This study aims to investigate the roles played by NaB and NaP in breast carcinogenesis by elucidating their potential anti-metastatic effects in the context of tumor migration, invasion, and EMT regulation in two distinct breast cancer cell lines, MCF-7 and MDA-MB-231.
The cytotoxic effect of both compounds on 3D spheroid formation was evaluated using a hanging drop assay. The anti-migratory and anti-invasive potentials of NaB and NaP were investigated through transwell migration and invasion assays. Moreover, their role in regulating epithelial-to-mesenchymal transition (EMT) was examined by assessing E-cadherin, vimentin, and β-catenin mRNA and protein expression levels through RT-qPCR and Western blot or flow cytometry. β-Catenin localization upon treatment was further visualized via immunofluorescence. Protein expression of MEK, p-MEK, ERK, and p-ERK was analyzed by Western blot.
Our results revealed a dose- and time-dependent impairment of spheroid formation in both cell lines, with NaB exerting a more potent effect than NaP. Both SCFAs were able to significantly inhibit migration and invasion of MDA-MB-231 cells following 24 h of treatment. Moreover, treatment with NaB or NaP altered the mRNA and protein profile of EMT-associated markers and abrogated the nuclear translocation of β-catenin. Finally, ERK and MEK phosphorylation was reduced in MDA-MB-231 and MCF-7 cells upon treatment with NaB, and less prominently with NaP.
Our study highlights the promising therapeutic potential of NaB and NaP, providing insight into their inhibitory effects on 3D formation, migration, and invasion through EMT regulation and deactivation of MEK/ERK signaling in breast cancer.
本研究旨在通过阐明丁酸钠（NaB）和丙酸钠（NaP）在两种不同的乳腺癌细胞系MCF-7和MDA-MB-231的肿瘤迁移、侵袭和上皮-间质转化（EMT）调节背景下的潜在抗转移作用，来研究它们在乳腺癌发生中的作用。
使用悬滴法评估两种化合物对三维球体形成的细胞毒性作用。通过Transwell迁移和侵袭试验研究NaB和NaP的抗迁移和抗侵袭潜力。此外，通过实时定量聚合酶链反应（RT-qPCR）、蛋白质免疫印迹法或流式细胞术评估E-钙黏蛋白、波形蛋白和β-连环蛋白的mRNA和蛋白质表达水平，来研究它们在调节上皮-间质转化（EMT）中的作用。通过免疫荧光进一步观察处理后β-连环蛋白的定位。通过蛋白质免疫印迹法分析MEK、磷酸化MEK（p-MEK）、细胞外信号调节激酶（ERK）和磷酸化ERK（p-ERK）的蛋白质表达。
我们的结果显示，两种细胞系中球体形成均呈现剂量和时间依赖性损伤，其中NaB的作用比NaP更强。两种短链脂肪酸在处理24小时后均能显著抑制MDA-MB-231细胞的迁移和侵袭。此外，用NaB或NaP处理会改变EMT相关标志物的mRNA和蛋白质谱，并消除β-连环蛋白的核转位。最后，用NaB处理后，MDA-MB-231和MCF-7细胞中ERK和MEK的磷酸化水平降低，而用NaP处理时降低程度较小。
我们的研究突出了NaB和NaP有前景的治疗潜力，揭示了它们通过调节EMT以及使乳腺癌中的MEK/ERK信号失活，从而对三维球体形成、迁移和侵袭产生抑制作用。
