Journal of Shandong University (Health Sciences) ›› 2026, Vol. 64 ›› Issue (6): 1-12.doi: 10.6040/j.issn.1671-7554.0.2025.1227

• Preclinical Medicine •    

Under hypoxic conditions, prolyl hydroxylase domain-containing protein 2 regulates ubiquitin-specific protease 22 to participate in the invasive biological behavior of esophageal squamous cell carcinoma

WANG Shenghai1, QI Yuhao2, HUANG Yinghua2, GONG Shaojun3, LI Jun2,4   

  1. 1. Department of Pathology and Pathophysiology, Medical College of Qinghai University, Xining 810001, Qinghai, China;
    2. Department of Thoracic Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan 250021, Shandong, China;
    3. Department of Thoracic Surgery, The First Peoples Hospital of Xining, Xining 810000, Qinghai, China;
    4. Department of Thoracic Surgery, Qinghai Red Cross Hospital, Xining 810000, Qinghai, China
  • Published:2026-06-29

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Abstract: Objective To investigate the effects of prolyl hydroxylase domain-containing protein 2(PHD2)on the malignant biological behaviors of esophageal squamous cell carcinoma(ESCC)in a hypoxic microenvironment, and to elucidate its regulatory relationship with ubiquitin-specific protease 22(USP22). Methods The high-throughput public gene expression database, gene expression omnibus(GEO), was utilized to analyze the expression of PHD2 mRNA in ESCC tissues and normal tissues. Western blotting was performed to evaluate PHD2 expression levels in a panel of esophageal squamous cell carcinoma(ESCC)cell lines(KYSE-410, KYSE-30, KYSE-150, EC-9706 and TE-1), with normal immortalized human esophageal epithelial cells(HET-1A, immortalized by SV40)serving as the control. ESCC cell models with PHD2 silencing(knockdown)and overexpression were established via transfection. Under normoxic and hypoxic(1%O2)conditions, the proliferative capacity of ESCC cells was evaluated using the CCK-8 assay, while the migratory ability was assessed via wound healing and Transwell migration assays. Furthermore, the effects of PHD2 silencing and overexpression on USP22 expression levels were evaluated to validate their regulatory relationship. Finally, co-immunoprecipitation(Co-IP)assays were performed to demonstrate the interaction between PHD2 and USP22. Results Analysis of public gene expression databases revealed that PHD2 mRNA expression levels were higher in normal tissues than in ESCC tissues. Compared with ESCC cells, the expression of PHD2 was significantly higher in normal esophageal squamous epithelial cells(HET-1A)(P<0.05). Hypoxia promoted the proliferation and migration abilities of ESCC cells. Silencing of PHD2 enhanced the proliferation and migration of ESCC cells, whereas PHD2 overexpression suppressed these abilities(P<0.05). After 24 hours of hypoxia, PHD2 expression decreased while USP22 expression increased(P<0.05). Furthermore, PHD2 silencing upregulated the expression of USP22(P<0.05), while PHD2 overexpression downregulated USP22 expression(P<0.05). Finally, Co-IP assays confirmed the physical interaction between PHD2 and USP22. Conclusion PHD2 is expressed at low levels in both ESCC tissues and cell lines, and its overexpression suppresses the proliferation and migration of ESCC cells. Under normoxic conditions, PHD2 overexpression downregulates USP22 expression; notably, under hypoxic conditions, PHD2 maintains its regulatory effect on USP22. This suggests that a regulatory relationship likewise exists between USP22 and PHD2 under the hypoxic conditions of solid tumors, and they synergistically contribute to the aggressive biological behaviors of ESCC. PHD2 may therefore serve as a novel therapeutic target for ESCC.

Key words: Prolyl hydroxylase domain-containing protein 2, Ubiquitin-specific protease 22, Esophageal squamous cell carcinoma, Proliferation, Migration

CLC Number: 

  • R735
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