Cytoglobin dependent transcriptomic changes determine sensitivity of melanoma to ferroptopsis. Cytoglobin dependent transcriptomic changes determine sensitivity of melanoma to ferroptopsis

In this study, we investigated the potential effect of CYGB on the cellular sensitivity towards (1S, 3R)-RAS-selective lethal small molecule (RSL3)-mediated ferroptosis in the G361 melanoma cells with abundant endogenous expression. Our findings show that an increased basal ROS level and higher degree of lipid peroxidation upon RSL3 treatment contributes to the increased sensitivity of CYGB knockdown G361 cells to ferroptosis. Furthermore, transcriptome analysis demonstrates the enrichment of multiple cancer malignancy pathways upon CYGB knockdown, supporting a tumor suppressive role for CYGB. Remarkably, CYGB knockdown also triggered activation of the NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome and subsequent induction of pyroptosis target genes. Altogether, we show that silencing of CYGB expression modulates cancer therapy sensitivity via modulation of ferroptosis and pyroptosis cell death signalling pathways. Overall design: Differential gene expression analysis of CYGB knockdown G361 melanoma cells under basal conditions (untreated) and upon treatment with ferroptosis inducer RSL3.

本研究以高表达内源性CYGB的G361黑色素瘤细胞为模型，探究了CYGB对(1S,3R)-RAS选择性致死小分子(RSL3)介导的铁死亡(ferroptosis)细胞敏感性的潜在调控效应。研究结果显示，经RSL3处理后，CYGB敲低的G361细胞基础活性氧(Reactive Oxygen Species, ROS)水平升高、脂质过氧化程度加剧，这与其对铁死亡的敏感性增强密切相关。进一步的转录组分析表明，CYGB敲低后多条肿瘤恶性相关通路发生富集，佐证了CYGB的肿瘤抑制功能。值得注意的是，CYGB敲低还激活了含NOD、LRR和PYRIN结构域的蛋白3(NLRP3)炎症小体，并诱导了焦亡(pyroptosis)相关靶基因的表达。综上，本研究证实，沉默CYGB的表达可通过调控铁死亡与焦亡细胞死亡信号通路，调节肿瘤治疗敏感性。整体实验设计：对处于基础培养状态（未处理）及经铁死亡诱导剂RSL3处理的CYGB敲低G361黑色素瘤细胞进行差异基因表达分析。