ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression

• DOI: 10.1186/s12964-025-02182-3
• Publication Date: 2025-04-28T07:48:50Z
• Authors: Mi Kyung Park; Hye Ja Lee; Jee Young Sung; Hyun Jung Byun; Hyun Ji Kim; Eun Ji Kim; Tuan Minh Nguyen; Gyeoung Jin Kang; Seung Hyun Oh; Jae Gal Shim; Ho Lee; Ki Taek Nam; Yong Yun Kim; Seung Bae Rho; Sang Gun Kim; Chang Hoon Lee
• Keywords: Expression (computer science)

Pancreatic cancer is the fourth leading cause of cancer-related deaths. Epithelial-mesenchymal transition (EMT) drives aggressive behaviour and unfavourable outcomes in this disease. The zinc finger E-box-binding homeobox 1 (ZEB1) transcription factor pivotal orchestrating EMT, promoting tumor cell mobility, metastasis, immune evasion through phosphorylation events. However, precise mechanisms underlying individual sites their relationship with ZEB1's functions vivo remain inadequately understood. We assessed EMT using various techniques, including reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunoblotting, microscopy, migration, invasion assays. ZEB1 knockdown was achieved via short hairpin RNA (shRNA), while plasmid transfection facilitated overexpression ZEB1, extracellular signal-regulated kinase (ERK1), 2 (ERK2). Co-immunoprecipitation assays were used to examine interaction between ERK1/2. PANC-1 HPAC cells transplanted an orthotopic mouse model for analysis. Sphingosylphosphorylcholine (SPC) induced a dose- time-dependent manner nuclear translocation ZEB1. Notably, ERK2 interacted catalysed serine 322 (S322) within molecule. Disrupting S322 hindered translocation, reduced programmed death-ligand (PD-L1) expression suppressed migration pancreatic cells. Furthermore, model, implantation phosphorylation-deficient (shZEB1/S322A) tumour formation metastasis. developed phosphoS322 detection antibody, which validated tissue samples from patients cancer. SPC induces phosphorylation, ERK2, rather than ERK1, targeting site. Inhibiting mitigates PD-L1 expression, progression antibody might be valuable tool predicting prognosis. These findings implicate as potential therapeutic target highlight phosphoZEB1 prognostic marker.