Timosaponin AIII inhibits gastric cancer by causing oxidative stress and blocking autophagic flux

IF 5 2区医学 Q2 Medicine

Translational Oncology Pub Date : 2025-07-29 DOI:10.1016/j.tranon.2025.102481

Chunyang Zhu , Shuming Chen , Yangyang Lu, Jialin Song, Shasha Wang, Jing Guo, Xiaoxi Han, YuanYuan Fang, Siyi Zhang, Wensheng Qiu, Weiwei Qi

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引用次数: 0

Abstract

Background

: Gastric cancer (GC) is a prevalent malignant tumor worldwide, with limited treatment targets. Timosaponin AIII (Tim AIII) is the naturally steroid saponin isolated from Anemarrhena, while this study initially confirms the anti-GC effect of Tim AIII.

Methods

: MTT assay, cell cycle analysis, and wound healing assay were used to evaluate the inhibitory effects of Tim AIII on GC cells (AGS and HGC27). Evaluate the oxidative stress (OS) by measuring reactive oxygen species (ROS) and malondialdehyde (MDA), as well as the Kelch-like ECH-associated protein 1 (Keap1) - Nuclear factor erythroid-derived 2-like 2 (Nrf2) pathway. RNA sequencing and proteomics analysis were utilized to investigate deeper molecular mechanisms. To track the autophagic flux using transmission electron microscope, detecting changes in autophagy-related pathway proteins, staining with LC3B and lysosome. Experiments related to cell viability, OS, and autophagy levels were performed on normal gastric mucosal epithelial cells (GES-1) as parallel controls. Finally, Nude mouse subcutaneous tumor model to evaluate the anti-GC ability in vivo.

Results

: Tim AIII inhibits the viability, proliferation, and migration of GC cells. Tim AIII causes OS in GC cells by the increasing intracellular ROS and MDA levels and inhibiting the Keap1-Nrf2 pathway. RNA sequencing and proteomics analysis mainly focused on the autophagy-associated pathways and lysosome in GC cells. Tim AIII activates autophagy, as indicated by an increase in the number of autophagosomes, inhibition of the PI3K-AKT pathway, and activation of the AMPK pathway in GC cells. However, Tim AIII inhibits autophagy-lysosome fusion and impairs lysosomal function, which results in autophagic flux blockage in GC cells. The Tim AIII concentration that significantly inhibited GC cells in this study was applied to GES-1 cells. The results showed that at this concentration, Tim AIII exhibited no significant cytotoxic effects on GES-1 cells, did not induce OS, and had no impact on autophagy. Finally, Tim AIII also has the ability to inhibit tumor growth in vivo.

Significance

: In summary, the results of our study indicate Tim AIII as a novel late-stage autophagy inhibitor, which may provide novel medical possibilities for GC.

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Timosaponin AIII通过引起氧化应激和阻断自噬通量抑制胃癌

背景：胃癌是一种世界范围内普遍存在的恶性肿瘤，其治疗靶点有限。Timosaponin AIII （Tim AIII）是一种天然甾类皂苷，从赤霉素中分离得到，本研究初步证实了Tim AIII的抗gc作用。方法：采用MTT法、细胞周期法、创面愈合法评价Tim AIII对胃癌细胞（AGS和HGC27）的抑制作用。通过测定活性氧（ROS）和丙二醛（MDA），以及Kelch-like ECH-associated protein 1 (Keap1) - Nuclear factor erythroid-derived 2-like 2 （Nrf2）通路来评估氧化应激（OS）。RNA测序和蛋白质组学分析用于研究更深层次的分子机制。透射电镜追踪自噬通量，检测自噬相关途径蛋白的变化，LC3B和溶酶体染色。以正常胃粘膜上皮细胞（GES-1）作为平行对照，进行细胞活力、OS和自噬水平相关的实验。最后建立裸鼠皮下肿瘤模型，在体内评价其抗gc能力。结果：Tim AIII抑制胃癌细胞的活力、增殖和迁移。Tim AIII通过增加细胞内ROS和MDA水平，抑制Keap1-Nrf2通路，导致GC细胞发生OS。RNA测序和蛋白质组学分析主要集中在GC细胞的自噬相关途径和溶酶体。在GC细胞中，Tim AIII可以激活自噬，增加自噬体的数量，抑制PI3K-AKT通路，激活AMPK通路。然而，Tim AIII抑制自噬-溶酶体融合，损害溶酶体功能，导致GC细胞自噬通量阻滞。将本研究中显著抑制GC细胞的Tim AIII浓度应用于GES-1细胞。结果表明，在该浓度下，Tim AIII对GES-1细胞无明显的细胞毒作用，不诱导OS，对自噬无影响。最后，Tim AIII在体内也具有抑制肿瘤生长的能力。意义：综上所述，我们的研究结果提示Tim AIII作为一种新型的晚期自噬抑制剂，可能为胃癌的治疗提供新的医学可能性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Translational Oncology ONCOLOGY-

CiteScore

8.40

自引率

2.00%

发文量

314

审稿时长

54 days

期刊介绍： Translational Oncology publishes the results of novel research investigations which bridge the laboratory and clinical settings including risk assessment, cellular and molecular characterization, prevention, detection, diagnosis and treatment of human cancers with the overall goal of improving the clinical care of oncology patients. Translational Oncology will publish laboratory studies of novel therapeutic interventions as well as clinical trials which evaluate new treatment paradigms for cancer. Peer reviewed manuscript types include Original Reports, Reviews and Editorials.