Jiangxi Medical College, Nanchang University, Nanchang, 330036, Jiangxi, China.
Jiangxi Provincial Key Laboratory of Tumor Metastasis and Precision Therapy, Nanchang, Jiangxi, China.
Sci Rep. 2024 Mar 18;14(1):6504. doi: 10.1038/s41598-024-57196-7.
This study aimed to investigate the role of autophagy, ferroptosis, and pyroptosis in the antitumour mechanism of harmine (Har) and its crosstalk in ovarian cancer. By transmission electron microscopy, we found that compared with those in the control group, the cytoplasm of human ovarian cancer cells (SKOV3) treated with Har showed increased numbers of autophagic vesicles, decreased intracellular mitochondrial volume, increased bilayer membrane density, and decreased cristae. Western blot, immunofluorescence, and monodasylcadaverine (MDC) staining all suggested that Har promoted autophagy in SKOV3 cells. LY294002 and siFOXO3 rescued the inhibition of the PI3K/AKT/mTOR/FOXO3 signalling pathway and the promotion of autophagy by Har. Additionally, the levels of ferroptosis- and pyroptosis-related proteins and the levels of Fe , glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) suggested that Har promoted ferroptosis and pyroptosis in SKOV3 cells. Interestingly, pretreatment with chloroquine (CQ), erastin, rapamycin (Rap), or ferrostatin-1 (Fer-1) increased or reversed the ferroptosis and pyroptosis promoted by Har, respectively. In vivo, the volume of tumours in the Har group was decreased, and immunohistochemistry revealed decreased levels of Ki-67 and GPX4 and increased levels of ATG5 and NARL3. In conclusion, Har exerts its anti-ovarian cancer effect not only by promoting autophagy by regulating the PI3K/AKT/mTOR/FOXO3 signalling pathway but also by promoting ferroptosis and pyroptosis. Additionally, there is complex crosstalk between autophagy, ferroptosis, and pyroptosis in ovarian cancer.
本研究旨在探讨自噬、铁死亡和细胞焦亡在哈尔滨（Har）抗卵巢癌作用机制及其相互作用中的作用。通过透射电子显微镜观察，我们发现与对照组相比，用 Har 处理的人卵巢癌细胞（SKOV3）的细胞质中自噬小体数量增加，细胞内线粒体体积减少，双层膜密度增加，嵴减少。Western blot、免疫荧光和单甲基二氢吖啶（MDC）染色均表明 Har 促进了 SKOV3 细胞的自噬。LY294002 和 siFOXO3 挽救了 Har 对 PI3K/AKT/mTOR/FOXO3 信号通路的抑制和自噬的促进作用。此外，铁死亡和细胞焦亡相关蛋白的水平以及 Fe、谷胱甘肽 (GSH)、丙二醛 (MDA) 和超氧化物歧化酶 (SOD) 的水平表明 Har 促进了 SKOV3 细胞的铁死亡和细胞焦亡。有趣的是，用氯喹 (CQ)、erastin、雷帕霉素 (Rap) 或 ferrostatin-1 (Fer-1) 预处理分别增加或逆转了 Har 促进的铁死亡和细胞焦亡。在体内，Har 组肿瘤体积减小，免疫组化显示 Ki-67 和 GPX4 水平降低，ATG5 和 NARL3 水平升高。综上所述，Har 通过调节 PI3K/AKT/mTOR/FOXO3 信号通路促进自噬，还通过促进铁死亡和细胞焦亡发挥其抗卵巢癌作用。此外，在卵巢癌中自噬、铁死亡和细胞焦亡之间存在复杂的相互作用。
