Department of Pharmacy, The Teaching Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610072, P.R. China.
Medicine School of Medical and Life Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610072, P.R. China.
Mol Med Rep. 2020 Oct;22(4):2826-2832. doi: 10.3892/mmr.2020.11376. Epub 2020 Jul 28.
Erastin, a classical inducer of non‑apoptotic cell death, exerts cytotoxicity in several types of cancer cells, including gastric cancer cells, by depleting glutathione, which is a primary cellular antioxidant, thus causing reactive oxygen species (ROS) accumulation. Although numerous studies have focused on the non‑apoptotic cell death induced by erastin, whether erastin induces apoptosis remains unknown. The present study confirmed the cytotoxicity of erastin in HGC‑27 cells and used a 30% inhibitory concentration (IC30, approximately 6.23 µM) for further analysis. The cell cycle analysis revealed that 6.23 µM of erastin inhibited proliferation by blocking the cell cycle at the G1/G0 phase. Further analysis also showed that 6.23 µM of erastin clearly inhibited HGC‑27 malignant behaviors, including migration, invasion, colony formation and tumor formation in soft agar. The observation of ROS accumulation due to erastin treatment led to determination of the effects of erastin on mitochondrial function and, as expected, erastin treatment decreased transcriptional activity and ATP production in mitochondria and disrupted the mitochondrial potential; these effects were reversed by the addition of the ROS scavenger NAC. To evaluate the effect of erastin in inducing apoptosis, HGC‑27 cells were treated with 6.23 µM of erastin for 7 days and then analyzed. Evident apoptotic cell death was induced by erastin and this apoptosis was reversed by the addition of an apoptosis inhibitor (zVAD) or NAC but not by the addition of a ferroptosis inhibitor (ferrostatin‑1). Furthermore, the detection of caspase‑3 and poly (adenosine diphosphate‑ribose) polymerase (PARP) also confirmed that treatment with erastin promoted the cleavage of caspase‑3 and PARP, which are hallmarks of apoptosis. Taken together, the present study revealed that a low dose of erastin inhibited malignant behavior and induced apoptosis by causing mitochondrial dysfunction.
依拉司汀是一种经典的非凋亡细胞死亡诱导剂，通过耗尽谷胱甘肽（一种主要的细胞抗氧化剂）在包括胃癌细胞在内的几种类型的癌细胞中发挥细胞毒性作用，从而导致活性氧（ROS）积聚。尽管许多研究都集中在依拉司汀诱导的非凋亡性细胞死亡上，但依拉司汀是否诱导细胞凋亡尚不清楚。本研究证实了依拉司汀在 HGC-27 细胞中的细胞毒性，并使用 30%抑制浓度（IC30，约 6.23μM）进行进一步分析。细胞周期分析显示，6.23μM 的依拉司汀通过将细胞周期阻滞在 G1/G0 期来抑制增殖。进一步的分析还表明，6.23μM 的依拉司汀明显抑制了 HGC-27 细胞的恶性行为，包括迁移、侵袭、集落形成和软琼脂中的肿瘤形成。由于依拉司汀处理导致 ROS 积累的观察导致确定了依拉司汀对线粒体功能的影响，正如预期的那样，依拉司汀处理降低了线粒体的转录活性和 ATP 产生，并破坏了线粒体电位；这些作用可以通过添加 ROS 清除剂 NAC 来逆转。为了评估依拉司汀诱导细胞凋亡的效果，用 6.23μM 的依拉司汀处理 HGC-27 细胞 7 天，然后进行分析。依拉司汀诱导明显的凋亡细胞死亡，这种凋亡可以通过添加凋亡抑制剂（zVAD）或 NAC 逆转，但不能通过添加铁死亡抑制剂（ferrostatin-1）逆转。此外，caspase-3 和多聚（腺苷二磷酸核糖）聚合酶（PARP）的检测也证实，依拉司汀处理促进了 caspase-3 和 PARP 的裂解，这是凋亡的标志。综上所述，本研究表明，低剂量的依拉司汀通过引起线粒体功能障碍来抑制恶性行为并诱导细胞凋亡。
