Department of Respiratory Medicine, Kanazawa University, Ishikawa, Japan
Department of Respiratory Medicine, Kanazawa University, Ishikawa, Japan.
Anticancer Res. 2021 Jun;41(6):2963-2977. doi: 10.21873/anticanres.15078.
BACKGROUND/AIM: Non-small-cell lung cancer (NSCLC) remains a significant cause of death despite the recent introduction of several improved therapeutics. Pemetrexed disodium heptahydrate (pemetrexed) is currently available in combination with a platinum derivative for patients with advanced non-squamous NSCLC for first-line treatment, and as a single agent for second-line treatment. However, the mechanisms underlying its anticancer activities are still not well understood. In this study, we evaluated the growth inhibitory effects of pemetrexed on PC9 (EGFR exon 19 deletion) cells and elucidated the underlying molecular mechanisms.
PC9 cells were treated with pemetrexed and then assessed for the cell viability, morphological and nuclear changes, antigenic alterations, SA-β-gal staining, and changes in protein expression.
Pemetrexed reduced the cell viability of PC9 cells and initiated cell morphological changes in a concentration-dependent manner. Pemetrexed significantly induced G phase arrest in a dose-dependent manner. The results demonstrated that pemetrexed induced apoptosis in PC9 cells, a change coupled with an increase in reactive oxygen species and a decrease in mitochondrial membrane potential. Pemetrexed decreased Bcl-2 expression, while Bax expression was increased, and cytochrome c was released. Furthermore, the expression of extrinsic pathway proteins, e.g. Fas/FasL, DR4/TRAIL, and Fas-associated protein with death domain, was increased by pemetrexed, which then activated caspase-8, caspase-9, and caspase-3 and induced poly (ADP-ribose) polymerase proteolysis.
This study revealed the mechanisms by which pemetrexed works an anticancer drug in the treatment of NSCLC.
背景/目的：尽管最近引入了几种改进的治疗方法，非小细胞肺癌（NSCLC）仍然是导致死亡的重要原因。培美曲塞二钠七水合物（培美曲塞）目前可与铂衍生物联合用于治疗晚期非鳞状 NSCLC 患者的一线治疗，也可作为二线治疗的单一药物。然而，其抗癌活性的机制仍不清楚。在这项研究中，我们评估了培美曲塞对 PC9（EGFR 外显子 19 缺失）细胞的生长抑制作用，并阐明了其潜在的分子机制。
用培美曲塞处理 PC9 细胞，然后评估细胞活力、形态和核变化、抗原改变、SA-β-半乳糖苷染色和蛋白表达变化。
培美曲塞降低了 PC9 细胞的细胞活力，并以浓度依赖的方式启动细胞形态变化。培美曲塞以剂量依赖性方式显著诱导 G 期停滞。结果表明，培美曲塞诱导 PC9 细胞凋亡，这种变化伴随着活性氧的增加和线粒体膜电位的降低。培美曲塞降低了 Bcl-2 的表达，同时增加了 Bax 的表达，并释放了细胞色素 c。此外，培美曲塞增加了外源性途径蛋白的表达，如 Fas/FasL、DR4/TRAIL 和 Fas 相关死亡域蛋白，然后激活了 caspase-8、caspase-9 和 caspase-3，并诱导多聚（ADP-核糖）聚合酶的蛋白水解。
本研究揭示了培美曲塞在治疗 NSCLC 中作为抗癌药物的作用机制。
