糖酵解酶糖三磷酸脱氢酶（GAPDH）是最为突出的保守蛋白之一，在某些半定量测定中广泛用作内参。除了参与糖酵解外，GAPDH还参与多种与癌症相关的生物过程，并且据报道，在多种癌症类型中普遍发生异常调控。因此，亟需详细解析GAPDH在癌症生理过程中的作用。全癌症分析显示，GAPDH在大多数癌症类型中广泛高表达，并且具有高GAPDH表达的肿瘤组织患者预后差的特点。GAPDH的表达与免疫细胞浸润及免疫检查点的一致性暗示了GAPDH与肿瘤微环境以及肿瘤发展之间的一定关联。基因集富集分析表明，GAPDH可能对多个与癌症相关的重要途径和生物过程做出贡献。多组学分析和体外细胞实验揭示了GAPDH的过表达由DNA拷贝数扩增和启动子甲基化修饰调控。重要的是，转录因子forkhead box M1（FOXM1）能够调控GAPDH的表达，证实其为一个癌基因并在多种癌症类型中普遍高表达。半定量染色质免疫沉淀、定量PCR和双荧光素酶报告实验显示FOXM1主要结合两个癌细胞系的GAPDH启动子区域。本研究结果揭示了GAPDH在肿瘤发展中的作用，因此引起对这一重要分子的关注，并对其在癌症研究中作为内参基因的作用提出疑问。© 2023作者。

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is one of the most prominent housekeeping proteins and is widely used as an internal control in some semi-quantitative assays. In addition to glycolysis, GAPDH is involved in several cancer-related biological processes and has been reported to be commonly dysregulated in multiple cancer types. Therefore, its role in the physiological process of cancer needs to be urgently elucidated. Pan-cancer analysis indicated that GAPDH is ubiquitously highly expressed in most cancer types, and that patients with a high GAPDH expression of in tumor tissues have a poor prognosis. The concordance of GAPDH expression in tumors with the infiltration of immune cells and immune checkpoints implies a certain association between GAPDH and the tumor microenvironment as well as tumor development. Gene Set Enrichment Analysis revealed that GAPDH may contribute to multiple important cancer-related pathways and biological processes. Multi-omics analysis and in vitro cell experiments revealed that GAPDH overexpression is regulated by DNA copy number amplification and promoter methylation modification. Importantly, a transcription factor, forkhead box M1 (FOXM1), which is capable of regulating GAPDH expression, was also identified and was confirmed to be an oncogene and ubiquitously highly expressed in multiple cancer types. Semi-quantitative chromatin immunoprecipitation, quantitative PCR, and dual-luciferase assays showed that FOXM1 mainly binds to the promoter region of GAPDH in two cancer cell lines. The present findings revealed the implication of GAPDH in tumor development, thus bringing attention to this important molecule and casting doubts on its role as an internal reference gene in cancer studies.© 2023 The Author(s).