Borhan Aylar, Bagherlou Ali, Ghayour Mohammad B
Department of Biology, Faculty of Science, University of Mohaghegh Ardabili, Ardabil, Iran.
Department of Biophysics, Faculty of Advanced Technologies, University of Mohaghegh Ardabili, Namin, Iran.
Tissue Cell. 2025 Apr;93:102718. doi: 10.1016/j.tice.2024.102718. Epub 2025 Jan 3.
Carnosic acid (CA) has potential anti-cancer properties, but its effectiveness can be improved by combining it with Folic acid (FA). This research aimed to evaluate the impact of CA and CA-FA conjugate on breast cancer cell lines (MCF-7, MDA-MB-231, and MCA10).
The viability of the cell lines was measured using the MTT assay, and the IC₅₀ was determined to compare the cytotoxicity of CA and CA-FA. The process of programmed cell death was investigated by utilizing Annexin V/PI staining, measuring caspase-3/7 activity, and real-time PCR for apoptotic gene expression. Reactive oxygen species (ROS) were also assessed to determine the extent of oxidative stress.
CA significantly decreased the viability of MCF-7 and MDA-MB-231 cells depending on the dosage, with CA-FA exhibiting enhanced cytotoxicity, particularly in MDA-MB-231 cells. The evaluation of IC₅₀ confirmed that conjugation with FA reduced the IC₅₀ of CA. Apoptosis analysis demonstrated increased apoptosis rates in MCF-7 and MDA-MB-231 cells exposed to treatment with CA and CA-FA, while MCA10 cells showed minimal effects. Caspase-3/7 activity was notably higher in CA-FA-treated cells. Gene expression analysis revealed elevated pro-apoptotic gene activity and reduced anti-apoptotic gene activity, with CA-FA having a more pronounced effect. Cells subjected to CA-FA treatment exhibited a significant increase in ROS levels.
These findings suggest that CA conjugation with FA enhances its cytotoxic effects and promotes apoptosis through increased apoptosis and ROS production. The research emphasizes the promise of CA-FA as a focused treatment approach for aggressive forms of breast cancer, underscoring the need for additional exploration of its practical uses in clinical settings.
鼠尾草酸（CA）具有潜在的抗癌特性，但其有效性可通过与叶酸（FA）结合得到提高。本研究旨在评估CA及CA - FA共轭物对乳腺癌细胞系（MCF - 7、MDA - MB - 231和MCA10）的影响。
使用MTT法测定细胞系的活力，并测定IC₅₀以比较CA和CA - FA的细胞毒性。通过利用膜联蛋白V/PI染色、测量半胱天冬酶 - 3/7活性以及对凋亡基因表达进行实时PCR来研究程序性细胞死亡过程。还评估了活性氧（ROS）以确定氧化应激的程度。
CA根据剂量显著降低了MCF - 7和MDA - MB - 231细胞的活力，CA - FA表现出增强的细胞毒性，特别是在MDA - MB - 231细胞中。IC₅₀的评估证实与FA共轭降低了CA的IC₅₀。凋亡分析表明，接受CA和CA - FA处理的MCF - 7和MDA - MB - 231细胞的凋亡率增加，而MCA10细胞显示出最小的影响。在CA - FA处理的细胞中，半胱天冬酶 - 3/7活性显著更高。基因表达分析显示促凋亡基因活性升高，抗凋亡基因活性降低，CA - FA的作用更明显。接受CA - FA处理的细胞中ROS水平显著增加。
这些发现表明，CA与FA共轭增强了其细胞毒性作用，并通过增加凋亡和ROS产生促进凋亡。该研究强调了CA - FA作为侵袭性乳腺癌靶向治疗方法的前景，强调需要在临床环境中进一步探索其实际用途。
