Soares Bruno Cesar Costa, Khine Hnin Ei Ei, Sritularak Boonchoo, Chanvorachote Pithi, Alduina Rosa, Sungthong Rungroch, Chaotham Chatchai
Pharmaceutical Sciences and Technology Program, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Front Pharmacol. 2024 Apr 11;15:1361085. doi: 10.3389/fphar.2024.1361085. eCollection 2024.
The upgrade of natural products for cancer treatment is essential since current anticancer drugs still pose severe side effects. Cymensifin A (Cym A) isolated from an orchid Cymbidium ensifolium has shown its potential to induce the death of several cancer cells; however, its underlying molecular mechanisms are hitherto unknown. Here, we conducted a set of preliminary tests to assess the cytotoxic effects of Cym A on non-small-cell lung cancer (NSCLC) cells (A549, H23, H292, and H460). A flow cytometry system and Western blot analyses were employed to unveil molecular mechanisms underlying cancer cell apoptosis caused by Cym A. Cym A at 25-50 μM caused the death of all NSCLC cells tested, and its cytotoxicity was comparable to cisplatin, a currently used anticancer drug. The compound induced apoptosis of all NSCLC cells in a dose-dependent manner (5-50 μM), proven by flow cytometry, but H460 cells showed more resistance compared to other cells tested. Cym A-treated H460 cells demonstrated increased reactive oxygen species (ROS) and downregulated antioxidants (catalase, superoxide dismutase, and thioredoxin). The compound also upregulated the tumor suppressor P53 and the pro-apoptotic protein BAX but downregulated pro-survival proteins (BCL-2 and MCL-1) and deactivated survival signals (AKT and ERK) in H460 cells. Cym A was proven to trigger cellular ROS formation, but P53 and BAX were 2-fold more activated by Cym A compared to those treated with hydrogen peroxide. Our findings also supported that Cym A exerted its roles in the downregulation of nuclear factor erythroid 2-related factor 2 (a regulator of cellular antioxidant activity) and the increased levels of cleaved poly (ADP-ribose) polymerase (PARP) and cleaved caspase 3/7 during apoptosis. We propose that Cym A induces lung cancer cell death via ROS-mediated apoptosis, while the modulation of cellular ROS/antioxidant activity, the upregulation of P53 and BAX, the downregulation or deactivation of BCL-2, MCL-1, AKT, and ERK, and the increased cleavage of PARP and caspase 3/7, were the elucidated underlying molecular mechanisms of this phytochemical. The compound can be a promising candidate for future anticancer drug development.
由于目前的抗癌药物仍有严重的副作用，因此升级天然产物用于癌症治疗至关重要。从兰花建兰中分离出的西孟西芬A（Cym A）已显示出诱导多种癌细胞死亡的潜力；然而，其潜在的分子机制迄今尚不清楚。在此，我们进行了一系列初步试验，以评估Cym A对非小细胞肺癌（NSCLC）细胞（A549、H23、H292和H460）的细胞毒性作用。采用流式细胞术系统和蛋白质印迹分析来揭示Cym A导致癌细胞凋亡的分子机制。25-50μM的Cym A导致所有测试的NSCLC细胞死亡，其细胞毒性与目前使用的抗癌药物顺铂相当。该化合物以剂量依赖性方式（5-50μM）诱导所有NSCLC细胞凋亡，流式细胞术已证实这一点，但与其他测试细胞相比，H460细胞表现出更强的抗性。经Cym A处理的H460细胞表现出活性氧（ROS）增加，抗氧化剂（过氧化氢酶、超氧化物歧化酶和硫氧还蛋白）下调。该化合物还上调了肿瘤抑制因子P53和促凋亡蛋白BAX，但下调了H460细胞中的促生存蛋白（BCL-2和MCL-1）并使生存信号（AKT和ERK）失活。已证实Cym A可触发细胞ROS形成，但与用过氧化氢处理的细胞相比，Cym A使P53和BAX的激活程度高2倍。我们的研究结果还支持，Cym A在下调核因子红细胞2相关因子2（细胞抗氧化活性的调节剂）以及在细胞凋亡期间增加裂解的聚（ADP-核糖）聚合酶（PARP）和裂解的半胱天冬酶3/7水平方面发挥作用。我们提出，Cym A通过ROS介导的凋亡诱导肺癌细胞死亡，而细胞ROS/抗氧化活性的调节、P53和BAX的上调、BCL-2、MCL-1、AKT和ERK的下调或失活以及PARP和半胱天冬酶3/7裂解的增加，是这种植物化学物质已阐明的潜在分子机制。该化合物可能是未来抗癌药物开发的有前途的候选物。
