铜死亡相关基因SLC31A1在肾细胞癌中的功能及预后价值

Functional and prognostic value of cuproptosis-related gene SLC31A1 in renal cell carcinoma

目的:探讨铜死亡相关基因SLC31A1在肾细胞癌的表达水平、肿瘤浸润免疫细胞相关性及其对预后的影响,并在体外实验中验证SLC31A1在肾细胞癌中的功能。方法:采用TCGA数据库收集肾细胞癌的临床数据及铜死亡相关基因转录组数据;采用HPA数据库收集蛋白表达数据;采用ssGSEA和Spearman分析计算免疫细胞浸润水平及其与铜死亡相关基因的关系;采用R语言比较肾细胞癌患者不同临床分期中SLC31A1的mRNA表达水平差异;采用Kaplan-Meier(KM)分析探索SLC31A1对肾细胞癌预后的影响;采用COX分析判断SLC31A1是否是独立预后因素;采用RT-qPCR检测SLC31A1 mRNA的表达水平。过表达质粒调控SLC31A1在细胞中的表达后采用克隆形成及划痕实验检测调控SLC31A1后对细胞增殖侵袭能力的影响。结果:铜死亡相关基因FDX1、DLD、DBT、SLC31A1、ATP7A、GCSH、DLST、DLAT、PDHA1、PDHB在肾细胞癌中的mRNA和蛋白水平均低于正常组织(P<0.05)。SLC31A1与17种瘤内免疫浸润细胞正相关(P<0.05)。SLC31A1与T分期、M分期和TNM分期负相关(P<0.05)。SLC31A1高表达的肾细胞癌患者的PFS和OS均优于SLC31A1低表达的患者(P<0.05)。单因素与多因素COX分析均显示SLC31A1是肾细胞癌的独立预后因素(P<0.05)。过表达SLC31A1后抑制了769-P细胞的增殖侵袭能力(P<0.05)。结论:铜死亡相关基因SLC31A1不仅与肾细胞癌免疫微环境和患者预后具有相关性,且可能参与调控肾细胞癌的增殖侵袭。

Objective:To explore the expression level of copper death related gene SLC31A1 in renal cell carcinoma(RCC),the correlation between tumor infiltration and immune cells,and its impact on prognosis,and further validate the function of SLC31A1 in RCC in vitro cell experiments.Methods:Clinical and transcriptome data of cuproptosis-related genes in RCC were collected from the TCGA database,while protein expression data were obtained from the HPA database.ssGSEA and Spearman's analysis was used to calculate the immune cell infiltration levels and their correlation with cuproptosis-related genes.The R language was used to compare the differences in SLC31A1 mRNA expression among RCC patients at different clinical stages Kaplan-Meier(KM)analysis was employed to assess the impact of SLC31A1 on RCC prognosis.Cox analysis was conducted to determine whether SLC31A1 was an independent prognostic factor.RT-qPCR was used to detect the expression level of the SLC31A1.Overexpression plasmids were used to regulate the expression of SLC31A1 in cells.Clone formation and wound-healing assays were used to detect the proliferation and invasion ability of regulated cells.Results:Cuproptosis-related genes FDX1,DLD,DBT,SLC31A1,ATP7A,GCSH,DLST,DLAT,PDHA1,and PDHB were found to be downregulated at the mRNA and protein levels in RCC compared to normal tissues(P<0.05).SLC31A1 exhibited a positive correlation with 17 types of tumor-infiltrating immune cells(P<0.05).SLC31A1 showed a negative correlation with the T stage,M stage,and TNM stage(P<0.05).RCC patients with high expression of SLC31A1 had better progression-free survival(PFS)and overall survival(OS)than low expression(P<0.05).Univariate and multivariate Cox analyses demonstrated that SLC31A1 was an independent prognostic factor for RCC(P<0.05).Overexpression of SLC31A1 inhibited the proliferative and invasive abilities of 769-P cells(P<0.05).Conclusion:SLC31A1 is not only associated with the immune microenvironment and prognosis of RCC,but also potentially involved in regulating RCC proliferation and invasion.