Jahani Mozhgan, Yarani Reza, Rezazadeh Davood, Tahmasebi Hadis, Hoseinkhani Zohreh, Kiani Sara, Mansouri Kamran
Department of Tissue Engineering & Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
Steno Diabetes Center Copenhagen, The Capital Region of Denmark Pediatrics, Herlev and Gentofte Hospital, Copenhagen University Hospital, Copenhagen, Denmark.
Curr Cancer Drug Targets. 2025;25(3):257-269. doi: 10.2174/0115680096288665240315072646.
Doxorubicin (DOX) is a chemotherapy drug that is widely used in cancer therapy, especially in Triple-Negative Breast Cancer (TNBC) patients. Nevertheless, cytoprotective autophagy induction by DOX limits its cytotoxic effect and drug resistance induction in patients. Therefore, finding a new way is essential for increasing the effectiveness of this drug for cancer treatment.
This study aimed to investigate the effect of L-lysine on DOX cytotoxicity, probably through autophagy modulation in TNBC cell lines.
We used two TNBC cell lines, MDA-MB-231 and MDA-MB-468, with various levels of autophagy activity. Cell viability after treatment with L-lysine alone and in combination therapy was evaluated by MTT assay. Reactive Oxygen Species (ROS), nitric oxide (NO) concentration, and arginase activity were assessed using flow cytometric analysis, Griess reaction, and arginase activity assay kit, respectively. Real-time PCR and western blot analysis were used to evaluate the L-lysine effect on the autophagy-related genes and protein expression. Cell cycle profile and apoptotic assay were performed using flow cytometric analysis.
The obtained data indicated that L-lysine in both concentrations of 24 and 32 mM increased the autophagy flux and enhanced the DOX cytotoxicity, especially in MDA-MB-231, which demonstrated higher autophagy activity than MDA-MB-468, by inducing ROS and NO production. Furthermore, L-lysine induced G2/M arrest autophagy cell death, while significant apoptotic changes were not observed.
These findings suggest that L-lysine can increase DOX cytotoxicity through autophagy modulation. Thus, L-lysine, in combination with DOX, may facilitate the development of novel adjunct therapy for cancer.
阿霉素（DOX）是一种化疗药物，广泛应用于癌症治疗，尤其是三阴性乳腺癌（TNBC）患者。然而，DOX诱导的细胞保护性自噬限制了其细胞毒性作用以及患者中的耐药性诱导。因此，寻找一种新方法对于提高该药物治疗癌症的有效性至关重要。
本研究旨在探讨L-赖氨酸对DOX细胞毒性的影响，可能是通过调节TNBC细胞系中的自噬来实现。
我们使用了两种具有不同自噬活性水平的TNBC细胞系，MDA-MB-231和MDA-MB-468。通过MTT法评估单独使用L-赖氨酸以及联合治疗后的细胞活力。分别使用流式细胞术分析、Griess反应和精氨酸酶活性测定试剂盒评估活性氧（ROS）、一氧化氮（NO）浓度和精氨酸酶活性。使用实时PCR和蛋白质印迹分析来评估L-赖氨酸对自噬相关基因和蛋白质表达的影响。使用流式细胞术分析进行细胞周期分析和凋亡检测。
获得的数据表明，24 mM和32 mM浓度的L-赖氨酸均增加了自噬通量并增强了DOX的细胞毒性，特别是在MDA-MB-231中，该细胞系表现出比MDA-MB-468更高的自噬活性，这是通过诱导ROS和NO的产生实现的。此外，L-赖氨酸诱导G2/M期阻滞的自噬性细胞死亡，而未观察到明显的凋亡变化。
这些发现表明L-赖氨酸可通过调节自噬增加DOX的细胞毒性。因此，L-赖氨酸与DOX联合使用可能有助于开发新型癌症辅助治疗方法。
