Zhang Xiwen, Liu Li, Chen Yanchen, Cheng Yumiao, Li Xinru, Pei Tianli, Zhang Fengmei
The Second Clinical Medical College of Henan University of Traditional Chinese Medicine, Zhengzhou, China.
Ophthalmology Department, Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou, China.
Transl Pediatr. 2025 Jul 31;14(7):1454-1470. doi: 10.21037/tp-2024-596. Epub 2025 Jul 28.
Disulfidoptosis, a newly recognised form of cell death, has been studied in many cancers but not in retinoblastoma. Therefore, this study aimed to investigate the role of disulfidoptosis-related genes (DRGs) in retinoblastoma.
Three retinoblastoma datasets (GSE208143, GSE97508, and GSE24673) were obtained from the Gene Expression Omnibus database. Differentially expressed DRGs (DE-DRGs) in retinoblastoma were identified, and key genes were screened using machine learning algorithms. Consensus cluster analysis was applied to identify the disulfidoptosis-based molecular subtypes. The pathways and immune characteristics of the subtypes were examined via gene set variation analysis, single-sample gene set enrichment analysis, and ESTIMATE. The expression of key genes was validated in retinoblastoma cells, and their roles were explored through functional experiments.
Compared with controls, the disulfidoptosis score was significantly lower (P<0.05) in retinoblastoma, and eight DE-DRGs were identified, suggesting the potential involvement disulfidoptosis in retinoblastoma. Consensus clustering revealed two molecular subtypes (C1 and C2). The C2 subtype exhibited multiple activated oncogenic pathways associated with tumourigenesis, lower infiltration levels of immune cells, lower immune and stromal scores, and reduced immune checkpoint expression. Least absolute shrinkage and selection operator and support vector machine-recursive feature elimination algorithms identified two key down-regulated genes-EPAS1 and SLC7A11-in retinoblastoma. The expression of EPAS1 and SLC7A11 was significantly correlated with oxidative phosphorylation and immune cell infiltration levels. EPAS1 overexpression significantly inhibited the viability, migration, and invasion of retinoblastoma cells and induced their apoptosis (P<0.05). Furthermore, EPAS1 overexpression significantly increased the relative nicotinamide adenine dinucleotide phosphate (NADP)+/reduced NADP (NADPH) ratio in retinoblastoma cells (P<0.05).
This study highlights the involvement of DRGs in retinoblastoma progression and molecular subtype heterogeneity. EPAS1 and SLC7A11 may serve as key targets for the diagnosis and treatment of retinoblastoma.
二硫键介导的细胞凋亡是一种新发现的细胞死亡形式，已在多种癌症中进行了研究，但尚未在视网膜母细胞瘤中开展研究。因此，本研究旨在探讨二硫键介导的细胞凋亡相关基因（DRGs）在视网膜母细胞瘤中的作用。
从基因表达综合数据库中获取了三个视网膜母细胞瘤数据集（GSE208143、GSE97508和GSE24673）。鉴定了视网膜母细胞瘤中差异表达的DRGs（DE-DRGs），并使用机器学习算法筛选关键基因。应用一致性聚类分析来识别基于二硫键介导的细胞凋亡的分子亚型。通过基因集变异分析、单样本基因集富集分析和ESTIMATE来研究亚型的通路和免疫特征。在视网膜母细胞瘤细胞中验证关键基因的表达，并通过功能实验探索其作用。
与对照组相比，视网膜母细胞瘤中的二硫键介导的细胞凋亡评分显著降低（P<0.05），并鉴定出8个DE-DRGs，提示二硫键介导的细胞凋亡可能参与视网膜母细胞瘤的发生发展。一致性聚类分析揭示了两种分子亚型（C1和C2）。C2亚型表现出与肿瘤发生相关的多种激活的致癌通路、免疫细胞浸润水平较低、免疫和基质评分较低以及免疫检查点表达降低。最小绝对收缩和选择算子以及支持向量机递归特征消除算法确定了视网膜母细胞瘤中两个关键的下调基因——EPAS1和SLC7A11。EPAS1和SLC7A11的表达与氧化磷酸化和免疫细胞浸润水平显著相关。EPAS1过表达显著抑制视网膜母细胞瘤细胞的活力、迁移和侵袭，并诱导其凋亡（P<0.05）。此外，EPAS1过表达显著提高了视网膜母细胞瘤细胞中烟酰胺腺嘌呤二核苷酸磷酸（NADP）+ /还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的相对比值（P<0.05）。
本研究强调了DRGs参与视网膜母细胞瘤的进展和分子亚型异质性。EPAS1和SLC7A11可能作为视网膜母细胞瘤诊断和治疗的关键靶点。
