亚硒酸钠通过铁死亡抑制KYSE-150食管鳞癌细胞增殖迁移活性

Sodium Selenite Inhibits the Proliferation and Migration of KYSE-150 Esophageal Squamous Cell Carcinoma Cells via Ferroptosis

摘要:

为研究亚硒酸钠（Na2SeO3）抑制KYSE-150食管鳞癌细胞的活性及机制。采用形态观察、细胞增殖抑制、划痕、Transwell等手段检测Na2SeO3对细胞增殖、迁移活性的影响；采用转录组测序确定该抑制发生的主要机制；分析Na2SeO3对细胞活性氧（Reactive Oxygen Species, ROS）、Fe2+、丙二醛（Malondialdehyde, MDA）及谷胱甘肽（Glutathione , GSH）含量的影响；透射电镜观察超微结构变化。结果显示，Na2SeO3有明显的抗KYSE-150细胞增殖及迁移活性：高浓度处理24 h增殖活性仅为对照组的50.89%；Transwell显示高浓度处理24 h，细胞穿过小室的数量为对照组的15.38%，划痕实验显示10 μmol·L-1组24 h的愈合速度为对照组的34.22%。测序结果表明，差异基因富集到铁死亡信号通路；细胞呈现铁依赖性ROS积累，高浓度组Fe2+/ROS分别为对照组的2.57/15.81倍；MDA含量升高但GSH含量下降，分别为对照组的8.93、0.39倍；亚细胞结构也呈现一定损伤，而Na2SeO3的抑癌过程可被Fer-1所缓解。综上，Na2SeO3可导致细胞内氧化还原系统失衡、亚细胞结构损伤，通过铁死亡方式抑制KYSE150细胞的增殖及迁移活性。本研究为预防和治疗肿瘤方面提供了应用前景。

Abstract:

To investigate the activity and mechanism of sodium selenite (Na?SeO?) in inhibiting the KYSE-150 esophageal squamous cell carcinoma cells. Morphological observation, cell proliferation inhibition assay, wound healing assay, and Transwell assay were employed to evaluate the inhibitory effects of Na?SeO? on KYSE-150 cell proliferation and migration. Transcriptome sequencing was performed to identify the main mechanism mediating this inhibition. The effects of Na?SeO? on the contents of Reactive Oxygen Species (ROS), Fe2?, Malondialdehyde (MDA), and Glutathione (GSH) in cells were analyzed, and the impact on subcellular structures was observed by transmission electron microscopy. The results showed that Na?SeO? exhibited significant activity in inhibiting the proliferation and migration of KYSE-150 cells. After 24 h of high-concentration treatment, the proliferative activity was only 50.89% of that in the control group. Transwell assay results indicated that the number of cells passing through the chamber in the high-concentration group at 24 h of treatment was 15.38% of that in the control group; wound healing assay showed that the healing rate of the 10 μmol·L?1 group at 24 h was 34.22% of that in the control group. Transcriptome sequencing results revealed that differentially expressed genes were enriched in the ferroptosis signaling pathway. Cells exhibited iron-dependent ROS accumulation, with Fe2? and ROS levels in the highest concentration group reaching 2.57-fold and 15.81-fold those of the control group, respectively. Additionally, MDA content was increased (8.93-fold of the control group), while GSH content was decreased (0.39-fold of the control group), and subcellular structures showed a certain degree of damage. Whereas the anticancer effect of Na2SeO3 can be attenuated by Fer-1. In conclusion, Na?SeO? can induce imbalance of the intracellular redox system and damage to subcellular structures, thereby inhibiting the proliferation and migration of KYSE-150 cells via the ferroptosis pathway. This study provides an application prospect for the prevention and treatment of tumors.