Hypoxia-induced PGK1 expression promotes esophageal squamous cell carcinoma progression via stimulating MYH9-mediated GSK3β/β-catenin signalling

Abstract

Background: Phosphoglycerate kinase 1 (PGK1) serves as a critical metabolic enzyme in the process of glycolysis and has many nonmetabolic functions in tumour progression. One of the most prevalent malignant tumours is still esophageal squamous cell carcinoma (ESCC), with high recurrence rates, high probabilities of metastasis, and poor prognoses. However, the molecular mechanisms and physiological contribution of PGK1 to ESCC carcinogenesis remain largely elusive.

Methods: Esophageal cancer bioinformatics analysis and tissue microarray analysis were employed to elucidate the aberrant expression of PGK1 during ESCC progression. The carcinogenic effect of PGK1 was examined using cell proliferation, migration and sphere formation assays. Mass spectrometry analysis, immunoprecipitation, ChIP and luciferase assays, hypoxia assays and in vitro and in vivo experiments were used to clarify the mechanism of the PGK1‒MYH9 interaction in the β-catenin/c-Myc signalling pathway.

Results: We clarified that in patients with ESCC, elevated PGK1 levels were linked to poor survival, tumour size, lymph node metastatic status, and TNM stage. In vivo and in vitro experimental analyses revealed that PGK1 promoted ESCC cell tumour stemness and EMT both in vivo and in vitro. Mechanistically, we discovered that PGK1 interacts with myosin-9 (MYH9), leading to MYH9-mediated ubiquitination-mediated degradation of GSK-3β, thereby triggering the β-catenin signalling pathway and transcriptionally increasing c-Myc expression. In addition, we found that hypoxic conditions upregulated PGK1, with HIF-1α transactivating PGK1 expression, further promoting the PGK1-MYH9 interaction and PGK1/MYH9/β-catenin/c-Myc axis activation.

Conclusions: PGK1 promotes ESCC tumourigenicity and migratory capacity by facilitating β-catenin-dependent c-Myc transcription. Under hypoxic conditions, the PGK1‒MYH9 interaction is strengthened, and HIF-1α-mediated transcription increases PGK1 expression, thereby activating the β-catenin/c-Myc signalling pathway. Taken together, PGK1 holds promise as a potential biomarker for predicting postoperative prognosis and recurrence in patients with ESCC.

Keywords

β-catenin signalling pathway / esophageal squamous cell carcinoma / hypoxia / MYH9 / PGK1

References

[1] Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024; 74(3): 229-263.

[2] Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. CA Cancer J Clin. 2023; 73(1): 17-48.

[3] Morgan E, Soerjomataram I, Rumgay H, et al. The global landscape of esophageal squamous cell carcinoma and esophageal adenocarcinoma incidence and mortality in 2020 and projections to 2040: new estimates from GLOBOCAN 2020. Gastroenterology. 2022; 163(3): 649-658.e642.

[4] Rumgay H, Arnold M, Laversanne M, et al. International trends in esophageal squamous cell carcinoma and adenocarcinoma incidence. Am J Gastroenterol. 2021; 116(5): 1072-1076.

[5] Zhang YQ, Chen T, Zhang C, et al. Endoscopic submucosal dissection for superficial proximal esophageal neoplasia is highly successful. Ann Surg. 2017; 266(6): 995-999.

[6] Luketich JD, Pennathur A, Awais O, et al. Outcomes after minimally invasive esophagectomy: review of over 1000 patients. Ann Surg. 2012; 256(1): 95-103.

[7] Mönig S, Chevallay M, Niclauss N, et al. Early esophageal cancer: the significance of surgery, endoscopy, and chemoradiation. Ann N Y Acad Sci. 2018; 1434(1): 115-123.

[8] Pennathur A, Farkas A, Krasinskas AM, et al. Esophagectomy for T1 esophageal cancer: outcomes in 100 patients and implications for endoscopic therapy. Ann Thorac Surg. 2009; 87(4): 1048-1054. discussion.

[9] Zhang K, Sun L, Kang Y. Regulation of phosphoglycerate kinase 1 and its critical role in cancer. Cell Commun Signal. 2023; 21(1): 240.

[10] Liu H, Wang X, Shen P, Ni Y, Han X. The basic functions of phosphoglycerate kinase 1 and its roles in cancer and other diseases. Eur J Pharmacol. 2022; 920: 174835.

[11] Hu H, Zhu W, Qin J, et al. Acetylation of PGK1 promotes liver cancer cell proliferation and tumorigenesis. Hepatology. 2017; 65(2): 515-528.

[12] He Y, Wang X, Lu W, et al. PGK1 contributes to tumorigenesis and sorafenib resistance of renal clear cell carcinoma via activating CXCR4/ERK signaling pathway and accelerating glycolysis. Cell Death Dis. 2022; 13(2): 118.

[13] Zhang Y, Yu G, Chu H, et al. Macrophage-associated PGK1 phosphorylation promotes aerobic glycolysis and tumorigenesis. Mol Cell. 2018; 71(2): 201-215.e207.

[14] Li X, Jiang Y, Meisenhelder J, et al. Mitochondria-translocated PGK1 functions as a protein kinase to coordinate glycolysis and the TCA cycle in tumorigenesis. Mol Cell. 2016; 61(5): 705-719.

[15] Liang C, Shi S, Qin Y, et al. Localisation of PGK1 determines metabolic phenotype to balance metastasis and proliferation in patients with SMAD4-negative pancreatic cancer. Gut. 2020; 69(5): 888-900.

[16] Li X, Qian X, Jiang H, et al. Nuclear PGK1 alleviates ADP-dependent inhibition of CDC7 to promote DNA replication. Mol Cell. 2018; 72(4): 650-660.e658.

[17] Hanahan D. Hallmarks of cancer: new dimensions. Cancer Discov. 2022; 12(1): 31-46.

[18] Chen Z, Han F, Du Y, Shi H, Zhou W. Hypoxic microenvironment in cancer: molecular mechanisms and therapeutic interventions. Signal Transduct Target Ther. 2023; 8(1): 70.

[19] Rey-Keim S, Schito L. Origins and molecular effects of hypoxia in cancer. Semin Cancer Biol. 2024; 106-107: 166-178.

[20] Wicks EE, Semenza GL. Hypoxia-inducible factors: cancer progression and clinical translation. J Clin Invest. 2022; 132(11): e159839.

[21] Hu Y, Smyth GK. ELDA: extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays. J Immunol Methods. 2009; 347(1-2): 70-78.

[22] He TC, Sparks AB, Rago C, et al. Identification of c-MYC as a target of the APC pathway. Science. 1998; 281(5382): 1509-1512.

[23] Weng AP, Millholland JM, Yashiro-Ohtani Y, et al. c-Myc is an important direct target of Notch1 in T-cell acute lymphoblastic leukemia/lymphoma. Genes Dev. 2006; 20(15): 2096-2109.

[24] Lin X, Li AM, Li YH, et al. Silencing MYH9 blocks HBx-induced GSK3β ubiquitination and degradation to inhibit tumor stemness in hepatocellular carcinoma. Signal Transduct Target Ther. 2020; 5(1): 13.

[25] Akanji MA, Rotimi D, Adeyemi OS. Hypoxia-inducible factors as an alternative source of treatment strategy for cancer. Oxid Med Cell Longev. 2019; 2019: 8547846.

[26] Zhan Q, Liu B, Situ X, et al. New insights into the correlations between circulating tumor cells and target organ metastasis. Signal Transduct Target Ther. 2023; 8(1): 465.

[27] Roy S, Sunkara RR, Parmar MY, Shaikh S, Waghmare SK. EMT imparts cancer stemness and plasticity: new perspectives and therapeutic potential. Front Biosci (Landmark Ed). 2021; 26(2): 238-265.

[28] Loh JJ, Ma S. Hallmarks of cancer stemness. Cell Stem Cell. 2024; 31(5): 617-639.

[29] Harada K, Pool Pizzi M, Baba H, Shanbhag ND, Song S, Ajani JA. Cancer stem cells in esophageal cancer and response to therapy. Cancer. 2018; 124(20): 3962-3964.

[30] Testa U, Castelli G, Pelosi E. Esophageal cancer: genomic and molecular characterization, stem cell compartment and clonal evolution. Medicines (Basel). 2017; 4(3): 67.

[31] Lv M, Gong Y, Liu X, et al. CDK7-YAP-LDHD axis promotes D-lactate elimination and ferroptosis defense to support cancer stem cell-like properties. Signal Transduct Target Ther. 2023; 8(1): 302.

[32] Pecci A, Ma X, Savoia A, Adelstein RS. MYH9: structure, functions and role of non-muscle myosin IIA in human disease. Gene. 2018; 664: 152-167.

[33] Godet I, Shin YJ, Ju JA, Ye IC, Wang G, Gilkes DM. Fate-mapping post-hypoxic tumor cells reveals a ROS-resistant phenotype that promotes metastasis. Nat Commun. 2019; 10(1): 4862.

[34] Bai R, Li Y, Jian L, Yang Y, Zhao L, Wei M. The hypoxia-driven crosstalk between tumor and tumor-associated macrophages: mechanisms and clinical treatment strategies. Mol Cancer. 2022; 21(1): 177.

[35] Guo Z, Zhang Y, Wang H, et al. Hypoxia-induced downregulation of PGK1 crotonylation promotes tumorigenesis by coordinating glycolysis and the TCA cycle. Nat Commun. 2024; 15(1): 6915.

[36] Qian X, Li X, Cai Q, et al. Phosphoglycerate kinase 1 phosphorylates Beclin1 to induce autophagy. Mol Cell. 2017; 65(5): 917-931.e916.

RIGHTS & PERMISSIONS

2025 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.