Abstract：
Protein disulfide isomerase A3 (PDIA3) is an endoplasmic reticulum (ER) protein. It has different functions including glycoprotein folding in the ER. The unfavorable prognosis of cancer patients was related to the abnormal PDIA3 expression level. However, it is unclear how PDIA3 correlates with the malignant characteristics of different tumors and its impact on tumor immunity. Pan-cancer data were downloaded from several databases for large-scale bioinformatics analysis. The immunological functions of PDIA3 were systematically explored at the single-cell sequencing level, including cell communication, cell metabolism, cell evolution and epigenetic modification. We performed immunofluorescence staining to visualize PDIA3 expression and infiltration of macrophages in pan-cancer samples. Further, we performed a loss-of-function assay of PDIA3 in vitro. The CCK8 assay, clone formation assay, and transwell assay were performed. M2 macrophages were co-cultured with different cell lines before the transwell assay was performed. The immunofluorescence staining of pan-cancer samples presented a higher expression of PDIA3 than those of the paired normal tissues. According to single-cell sequencing analysis, expression of PDIA3 was closely associated with cell communication, cell metabolism, cell evolution and epigenetic modification. The knockdown of PDIA3 in tumor cells inhibited cell proliferation and invasion, and restrained cocultured M2 macrophage migration. Furthermore, PDIA3 displayed predictive value in immunotherapy response in human cancer cohorts, indicating a potential therapeutic target. Our study showed that PDIA3 was associated with tumor malignant characteristics and could mediate the migration of M2 macrophages in various tumor types. PDIA3 could be a promising target to achieve tumor control and improve the immune response on a pan-cancer scale.
摘要：
蛋白二硫键异构酶A3(PDIA3)是一种内质网(ER)蛋白。它具有不同的功能,包括在ER中的糖蛋白折叠。肿瘤患者的不良预后与PDIA3表达水平异常有关。然而,目前尚不清楚PDIA3与不同肿瘤的恶性特征及其对肿瘤免疫的影响。从多个数据库下载泛癌症数据用于大规模生物信息学分析。在单细胞测序水平上系统地探讨了PDIA3的免疫学功能,包括细胞通信，细胞代谢，细胞进化和表观遗传修饰。我们进行了免疫荧光染色，以显示泛癌样品中PDIA3的表达和巨噬细胞的浸润。Further,我们在体外进行了PDIA3功能丧失试验.CCK8测定，克隆形成测定，并进行了transwell测定。在进行transwell测定之前,将M2巨噬细胞与不同细胞系共培养。泛癌样品的免疫荧光染色显示PDIA3的表达高于配对的正常组织。根据单细胞测序分析,PDIA3的表达与细胞通讯密切相关，细胞代谢，细胞进化和表观遗传修饰。PDIA3在肿瘤细胞中的敲低抑制细胞增殖和侵袭,并抑制共培养的M2巨噬细胞迁移。此外，PDIA3在人类癌症队列的免疫治疗反应中显示出预测价值，表明潜在的治疗目标。我们的研究表明，PDIA3与肿瘤的恶性特征有关，并且可以介导M2巨噬细胞在各种肿瘤类型中的迁移。PDIA3可能是在泛癌症规模上实现肿瘤控制和改善免疫应答的有希望的靶标。
