Table_1_Protein Disulfide-Isomerase A3 Is a Robust Prognostic Biomarker for Cancers and Predicts the Immunotherapy Response Effectively.xlsx

BackgroundProtein disulfide isomerase A3 (PDIA3) is a member of the protein disulfide isomerase (PDI) family that participates in protein folding through its protein disulfide isomerase function. It has been reported to regulate the progression of several cancers, but its function in cancer immunotherapy is unknown.MethodsThe RNA-seq data of cancer and normal tissues were downloaded from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) databases. The Cbioportal dataset was used to explore the genomic alteration information of PDIA3 in pan-cancer. Human Protein Atlas (HPA) and ComPPI websites were employed to mine the protein information of PDIA3, and western blot assay was performed to monitor the upregulated PDIA3 expression in clinical GBM samples. The univariate Cox regression and the Kaplan–Meier method were utilized to appraise the prognostic role of PDIA3 in pan-cancer. Gene Set Enrichment Analysis (GSEA) was applied to search the associated cancer hallmarks with PDIA3 expression. TIMER2.0 was the main platform to investigate the immune cell infiltrations related to PDIA3 in pan-cancer. The associations between PDIA3 and immunotherapy biomarkers were performed by Spearman correlation analysis. The immunoblot was used to quantify the PDIA3 expression levels, and the proliferative and invasive ability of glioma cells was determined by colony formation and transwell assays.FindingsPDIA3 is overexpressed in most cancer types and exhibits prognosis predictive ability in various cancers, and it is especially expressed in the malignant cells and monocytes/macrophages. In addition, PDIA3 is significantly correlated with immune-activated hallmarks, cancer immune cell infiltrations, and immunoregulators, and the most interesting finding is that PDIA3 could significantly predict anti-PDL1 therapy response. Besides, specific inhibitors that correlated with PDIA3 expression in different cancer types were also screened by using Connectivity Map (CMap). Finally, knockdown of PDIA3 significantly weakened the proliferative and invasive ability of glioma cells.InterpretationThe results revealed that PDIA3 acts as a robust tumor biomarker. Its function in protein disulfide linkage regulation could influence protein synthesis, degradation, and secretion, and then shapes the tumor microenvironment, which might be further applied to develop novel anticancer inhibitors.

背景：蛋白质二硫键异构酶A3（PDIA3）为蛋白质二硫键异构酶（PDI）家族成员之一，通过其蛋白质二硫键异构酶功能参与蛋白质折叠过程。据文献报道，PDIA3可调节多种癌症的进展，但其于癌症免疫治疗中的功能尚不明确。方法：从癌症基因组图谱（TCGA）和基因型-组织表达（GTEx）数据库中下载了癌症和正常组织的RNA测序数据。利用Cbioportal数据集探究了PDIA3在泛癌中的基因组变异信息。人类蛋白质图谱（HPA）和ComPPI网站被用于挖掘PDIA3的蛋白质信息，并通过西 Blot实验监测了临床胶质母细胞瘤（GBM）样本中PDIA3的上调表达。采用单变量Cox回归和Kaplan-Meier方法评估了PDIA3在泛癌中的预后作用。应用基因集富集分析（GSEA）以探寻与PDIA3表达相关的癌症标志。TIMER2.0平台为主要研究工具，探究了与PDIA3相关的免疫细胞浸润情况。通过Spearman相关性分析评估了PDIA3与免疫治疗生物标志物之间的关联。免疫印迹用于量化PDIA3的表达水平，而胶质瘤细胞的增殖和侵袭能力则通过集落形成和Transwell实验来确定。发现：PDIA3在大多数癌症类型中过表达，并在多种癌症中显示出预后预测能力，尤其是在恶性细胞和单核细胞/巨噬细胞中表达。此外，PDIA3与免疫激活标志、癌症免疫细胞浸润和免疫调节因子显著相关，最引人注目的是，PDIA3能够显著预测抗-PDL1治疗反应。此外，通过连接组学（CMap）筛选出与不同癌症类型中PDIA3表达相关的特定抑制剂。最终，敲低PDIA3显著削弱了胶质瘤细胞的增殖和侵袭能力。解释：研究结果揭示了PDIA3作为一项稳健的肿瘤生物标志物的作用。其在蛋白质二硫键连接调控中的作用可能影响蛋白质的合成、降解和分泌，进而塑造肿瘤微环境，这可能进一步应用于新型抗癌抑制剂的研发。