Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand.
Functional Proteomics Technology Laboratory, Functional Ingredients and Food Innovation Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology for Development Agency, Pathum Thani, Thailand.
PeerJ. 2023 Mar 10;11:e15049. doi: 10.7717/peerj.15049. eCollection 2023.
Menthol, a natural compound in peppermint leaves, has several biological activities, including antioxidant, anti-inflammatory, antiviral, antibacterial and anticancer properties. This study revealed the anti-leukemic effects and its underlying mechanisms of the menthol related apoptosis signaling pathway and autophagy in both NB4 and Molt-4 leukemic cell lines.
Both leukemic cells were treated with menthol in various concentration. Cell viability was assessed using MTT assay, whereas apoptosis and autophagy were analyzed by flow cytometry using Annexin V-FITC/PI and anti-LC3/FITC antibodies staining, respectively. Apoptotic and autophagic related gene and protein expression were detected using RT-qPCR and western blot analysis, respectively. Moreover, STITCH database was used to predicts the interaction between menthol and proposed proteins.
Menthol significantly decreased cell viability in NB4 and Molt-4 cell lines in dose dependent manner. In combination of menthol and daunorubicin, synergistic cytotoxic effects were observed in leukemic cells. However, there was a minimal effect found on normal, peripheral blood mononuclear cells (PBMCs). Moreover, menthol significantly induced apoptosis induction upregulation of caspase-3, BAX, p53 and downregulation of MDM2 mRNA expression. Autophagy was also induced by menthol through upregulating ATG3 and downregulating mTOR mRNA expression. For protein expression, menthol significantly increased caspase-3 whereas decreased mTOR in both leukemic cells. Conclusions. These results suggest that menthol exhibits cytotoxic activities by inhibition of cell proliferation, induction of apoptosis and autophagy through activating the caspase cascade, altering BAX and p53/MDM2, and regulating autophagy the ATG3/mTOR signaling pathway.
薄荷醇是薄荷叶中的一种天然化合物，具有多种生物学活性，包括抗氧化、抗炎、抗病毒、抗菌和抗癌特性。本研究揭示了薄荷醇相关凋亡信号通路和自噬在 NB4 和 Molt-4 白血病细胞系中的抗白血病作用及其潜在机制。
用不同浓度的薄荷醇处理两种白血病细胞。用 MTT 法评估细胞活力，用 Annexin V-FITC/PI 和抗 LC3/FITC 抗体染色通过流式细胞术分析凋亡和自噬，分别。用 RT-qPCR 和 Western blot 分析检测凋亡和自噬相关基因和蛋白表达。此外，还使用 STITCH 数据库预测薄荷醇与拟议蛋白之间的相互作用。
薄荷醇显著降低 NB4 和 Molt-4 细胞系中细胞活力呈剂量依赖性。薄荷醇与柔红霉素联合使用时，在白血病细胞中观察到协同细胞毒性作用。然而，在正常外周血单核细胞 (PBMCs) 中发现作用最小。此外，薄荷醇显著诱导凋亡诱导 caspase-3、BAX、p53 上调和 MDM2 mRNA 表达下调。自噬也通过上调 ATG3 和下调 mTOR mRNA 表达被薄荷醇诱导。对于蛋白表达，薄荷醇显著增加 caspase-3，而降低 mTOR 在两种白血病细胞中。结论。这些结果表明，薄荷醇通过激活 caspase 级联、改变 BAX 和 p53/MDM2 以及调节自噬，抑制细胞增殖、诱导凋亡和自噬，发挥细胞毒性作用。自噬通过激活 caspase 级联、改变 BAX 和 p53/MDM2 以及调节自噬，发挥细胞毒性作用。自噬通过激活 caspase 级联、改变 BAX 和 p53/MDM2 以及调节自噬，发挥细胞毒性作用。
