基于干性指数模型对肉瘤预后、免疫浸润和铁死亡调控的分析  

Analysis of regulation of prognosis,immune infiltration,and ferroptosis in sarcoma based on stemness index model

作　　者：魏婧娴 孟莲 孙皓 张田田 刘春霞 Wei Jingxian;Meng Lian;Sun Hao;Zhang Tiantian;Liu Chunxia(Department of Pathology,Shihezi University School of Medicine/Department of Pathology,The First Affiliated Hospital,Shihezi University,Shihezi 832000,Xinjiang Uygur Autonomous Region,China;Department of Pathology,The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,Guangdong Province,China)

机构地区：[1]石河子大学医学院病理学系/石河子大学第一附属医院病理科,新疆维吾尔自治区石河子市832000 [2]广州医科大学附属第二医院病理科,广东省广州市510260

出　　处：《中国组织工程研究》2025年第19期4151-4160,共10页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81960485),项目负责人:刘春霞;国家自然科学基金项目(82060487),项目负责人:孟莲;2022年广州市科技计划市校联合项目(202201020104),项目负责人:刘春霞。

摘　　要：背景:干性指数可能与肉瘤预后、免疫浸润有关,但具体调控机制与特征基因尚不清楚。目的:利用基因干性指数模型探究肉瘤干细胞与患者预后和免疫浸润的关系,并识别肉瘤干细胞铁死亡特征基因。方法:从癌症基因组图谱(TCGA)中获取肉瘤RNA测序数据及相关临床信息。利用肉瘤干性指数,对获取的肉瘤RNA测序数据进行分组。利用生存数据进行组间预后情况分析,获取组间差异表达基因后进行通路富集和免疫浸润分析。获得铁死亡相关差异基因,构建蛋白互作网络并进行预后相关性分析。培养横纹肌肉瘤细胞系,分为贴壁细胞组和干细胞组,贴壁细胞组不予任何干预,干细胞组通过无血清培养处理以诱导横纹肌肉瘤干细胞富集,qRT-PCR验证干细胞相关标志物、铁死亡相关差异基因以及铁死亡相关标志物的mRNA表达。结果与结论:①以干性指数中位数为界将患者分成高干性指数组和低干性指数组,通过患病风险预测显示高干性指数组患者的无进展生存期低于低干性指数组,提示预后不佳;②GO、KEGG分析显示肉瘤高、低干性指数组差异表达基因参与不同的细胞通路;高干性指数组和低干性指数组之间免疫浸润存在差异;差异基因中有23个铁死亡相关基因,其中9个可构成相关性较强的蛋白互作网络,其中IDO1、IFNG、AQP5高表达的肉瘤患者预后更好,CA9高表达的肉瘤患者预后不佳;③qRT-PCR结果显示,与贴壁细胞组相比,干细胞组干细胞相关标志物NANOG、SOX2和OCT4 mRNA表达显著升高(P<0.05);与贴壁细胞组相比,干细胞组铁死亡相关标志物SLC7A11的mRNA表达显著降低(P<0.05),ACSL4、GPX4、FTH1和COX2的mRNA表达显著升高(P<0.05);与贴壁细胞组相比,干细胞组差异基因CA9的mRNA表达显著降低,IDO1、IFNG、AQP5的mRNA表达显著升高(P<0.05)。经上述生物信息学分析与实验验证,干细胞与肉瘤生存、铁死亡过程密�BACKGROUND:The stemness index may be associated with the prognosis and immune infiltration of sarcoma,but the specific regulatory mechanism and characteristic genes have yet to be fully elucidated.OBJECTIVE:To investigate the correlation between stem cells and prognosis as well as immune infiltration in sarcoma employing the gene stemness index model and to identify the ferroptosis signature genes associated with sarcoma stem cells.METHODS:The sarcoma RNA sequencing data and related clinical information were obtained from the Cancer Genome Atlas(TCGA).The sarcoma RNA sequencing data were grouped using the sarcoma stemness index.Survival data were used to analyze prognosis between groups.Differentially expressed genes were obtained for pathway enrichment and immune infiltration analysis.Ferroptosis-related differential genes were used to construct a protein interaction network and analyze prognostic correlation.Rhabdomyosarcoma cell lines were cultured and divided into adherent cell group and stem cell group.The adherent cell group received no intervention,while the stem cell group was treated with serum-free culture to enrich stem cells in rhabdomyosarcoma cells.qRT-PCR was used to evaluate stemness markers,ferroptosis-related genes,and mRNA expression of ferroptosis-related markers in the cells.RESULTS AND CONCLUSION:(1)Patients were divided into high and low stemness index groups based on the median stemness index.The progressionfree survival of patients in the high stemness index group was lower than that in the low stemness index group by disease risk prediction,suggesting poor prognosis.(2)According to GO and KEGG analysis,the groups with high and low stemness indices differed from one another.There were differences in immune infiltration between the high and low stemness index groups.Nine of the 23 ferroptosis-related genes in the differential genes have the potential to establish a highly correlated network of protein interactions.Patients with high expression of IDO1,IFNG,and AQP5 have a better prognosis,

关 键 词：肉瘤 干细胞 干性指数 铁死亡 生存预后 免疫浸润 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R36