1. 土贝母苷乙通过诱导铁自噬抑制 非小细胞肺癌细胞增殖.
- Author
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杨翘伊, 张春云, 孙硕, 李文敏, 黄鑫, 梁艳, 张伟伟, 李怀永, and 杨清竹
- Abstract
AIM: This study aimed to explore the induction of ferroptosis in non-small-cell lung cancer (NSCLC) cells by tubeimoside II (TBMS II) and to elucidate the underlying molecular mechanisms. METHODS: H460 NSCLC cells were cultured in vitro. Cell survival rates were assessed by using MTT assays, and doses of TBMS II resulting in below 50% survival were selected for further experimentation. Cell migration was evaluated using Transwell assays and the effects of TBMS II on H460 cell proliferation were assessed by colony formation assays. Flow cytometry and fluorescence microscopy were used to assess changes in lipid peroxidation (lipid ROS), and the levels of GSH, T-AOC, MDA, and Fe2+ were measured using commercial kits. Protein levels of GPX4, SLC7A11, FTH1, NCOA4, P62, and LC3 were examined using Western blot. Changes in mitochondrial structure were detected by transmission electron microscopy, and immunofluorescence was used to assess LC3 co-localization of FTH1 and NCOA4, as well as co-localization of LC3 and NCOA4 with lysosomes. RESULTS: Compared with the control group, TBMS II dose-dependently reduced H460 cell viability, migration, and clone formation, accompanied by the appearance of vacuoles within the cells. TBMS II treatment also led to decreased GSH and T-AOC levels, while increasing the cellular contents of MDA, indicating oxidative stress. Additionally, there was a decrease in the expression of the antioxidant proteins SLC7A11 and GPX4 in the cells, while lipid ROS and Fe2+ levels were increased in proportion to the TBMS II concentration. The ferroptosis inhibitor ferrostatin-1 reversed cell death caused by TBMS II, suggesting ferroptosis induction. Furthermore, increasing the TBMS II concentration resulted in an upregulation of the autophagy marker proteins LC3 II/LC3 I and P62, indicative of increased autophagy. TBMS II also affected mitochondrial morphology in the cells, as seen in reduced mitochondrial fluorescence intensity. Protein expression of NCOA4 increased with higher TBMS II concentrations, while that of FTH1 decreased. Co-localization of LC3 II with FTH1 and NCOA4, as well as the lysosomal association of LC3 II and FTH1, also increased in a dosedependent manner. CONCLUSION: TBMS II induces ferritinophagy in H460 cells, leading to decreased cell viability and increased ferroptosis. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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