ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression
Abstract Background 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 is pivotal in orchestrating...
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BMC
2025-04-01
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| Series: | Cell Communication and Signaling |
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| Online Access: | https://doi.org/10.1186/s12964-025-02182-3 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Mi Kyung Park |
| collection | DOAJ |
| description | Abstract Background 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 is pivotal in orchestrating EMT, promoting tumor cell mobility, metastasis, and immune evasion through phosphorylation events. However, the precise mechanisms underlying individual phosphorylation sites and their relationship with ZEB1’s functions in vivo remain inadequately understood. Methods We assessed EMT using various techniques, including reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunoblotting, microscopy, migration, and invasion assays. ZEB1 knockdown was achieved via short hairpin RNA (shRNA), while plasmid transfection facilitated the overexpression of ZEB1, extracellular signal-regulated kinase 1 (ERK1), and extracellular signal-regulated kinase 2 (ERK2). Co-immunoprecipitation and kinase assays were used to examine the interaction between ZEB1 and ERK1/2. PANC-1 and HPAC cells were transplanted in an orthotopic mouse model for in vivo analysis. Results Sphingosylphosphorylcholine (SPC) induced EMT in PANC-1 and HPAC cells in a dose- and time-dependent manner through the phosphorylation and nuclear translocation of ZEB1. Notably, ERK2 interacted with ZEB1 and catalysed the phosphorylation of serine 322 (S322) within the ZEB1 molecule. Disrupting S322 phosphorylation hindered ZEB1’s nuclear translocation, leading to reduced programmed death-ligand 1 (PD-L1) expression and suppressed migration and invasion of pancreatic cancer cells. Furthermore, in an orthotopic mouse model, implantation of S322 phosphorylation-deficient (shZEB1/S322A) pancreatic cancer cells suppressed tumour formation and metastasis. We developed a phosphoS322 detection antibody, which validated ZEB1 phosphorylation in pancreatic cancer cells and tissue samples from patients with pancreatic cancer. Conclusion SPC induces ZEB1 phosphorylation, with ERK2, rather than ERK1, targeting the S322 site. Inhibiting S322 phosphorylation mitigates EMT, PD-L1 expression, and progression of pancreatic cancer. The phosphoS322 detection antibody might be a valuable tool for predicting pancreatic cancer prognosis. These findings implicate ERK2 as a potential therapeutic target for pancreatic cancer and highlight phosphoZEB1 as a valuable prognostic marker. |
| format | Article |
| id | doaj-art-ff37fa0771054c14b334910474fddda5 |
| institution | OA Journals |
| issn | 1478-811X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj-art-ff37fa0771054c14b334910474fddda52025-08-20T01:47:32ZengBMCCell Communication and Signaling1478-811X2025-04-0123111810.1186/s12964-025-02182-3ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progressionMi Kyung Park0Hye Ja Lee1Jee Young Sung2Hyun Jung Byun3Hyun Ji Kim4Eun Ji Kim5Tuan Minh Nguyen6Gyeoung Jin Kang7Seung Hyun Oh8Jae Gal Shim9Ho Lee10Ki Taek Nam11Yong Yun Kim12Seung Bae Rho13Sang Gun Kim14Chang Hoon Lee15Department of Biomedical Science, Hwasung Medi-Science UniversityBK21 FOUR Team and Integrated Research Institute for Drug Development, College of Pharmacy, Dongguk UniversityNational Cancer CenterBK21 FOUR Team and Integrated Research Institute for Drug Development, College of Pharmacy, Dongguk UniversityBK21 FOUR Team and Integrated Research Institute for Drug Development, College of Pharmacy, Dongguk UniversityBK21 FOUR Team and Integrated Research Institute for Drug Development, College of Pharmacy, Dongguk UniversityBK21 FOUR Team and Integrated Research Institute for Drug Development, College of Pharmacy, Dongguk UniversityLillehei Heart Institute, University of MinnesotaCollege of Veterinary Medicine, Seoul National UniversityNational Cancer CenterNational Cancer CenterSeverance Biomedical Science Institute, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of MedicineNational Cancer CenterNational Cancer CenterBK21 FOUR Team and Integrated Research Institute for Drug Development, College of Pharmacy, Dongguk UniversityBK21 FOUR Team and Integrated Research Institute for Drug Development, College of Pharmacy, Dongguk UniversityAbstract Background 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 is pivotal in orchestrating EMT, promoting tumor cell mobility, metastasis, and immune evasion through phosphorylation events. However, the precise mechanisms underlying individual phosphorylation sites and their relationship with ZEB1’s functions in vivo remain inadequately understood. Methods We assessed EMT using various techniques, including reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunoblotting, microscopy, migration, and invasion assays. ZEB1 knockdown was achieved via short hairpin RNA (shRNA), while plasmid transfection facilitated the overexpression of ZEB1, extracellular signal-regulated kinase 1 (ERK1), and extracellular signal-regulated kinase 2 (ERK2). Co-immunoprecipitation and kinase assays were used to examine the interaction between ZEB1 and ERK1/2. PANC-1 and HPAC cells were transplanted in an orthotopic mouse model for in vivo analysis. Results Sphingosylphosphorylcholine (SPC) induced EMT in PANC-1 and HPAC cells in a dose- and time-dependent manner through the phosphorylation and nuclear translocation of ZEB1. Notably, ERK2 interacted with ZEB1 and catalysed the phosphorylation of serine 322 (S322) within the ZEB1 molecule. Disrupting S322 phosphorylation hindered ZEB1’s nuclear translocation, leading to reduced programmed death-ligand 1 (PD-L1) expression and suppressed migration and invasion of pancreatic cancer cells. Furthermore, in an orthotopic mouse model, implantation of S322 phosphorylation-deficient (shZEB1/S322A) pancreatic cancer cells suppressed tumour formation and metastasis. We developed a phosphoS322 detection antibody, which validated ZEB1 phosphorylation in pancreatic cancer cells and tissue samples from patients with pancreatic cancer. Conclusion SPC induces ZEB1 phosphorylation, with ERK2, rather than ERK1, targeting the S322 site. Inhibiting S322 phosphorylation mitigates EMT, PD-L1 expression, and progression of pancreatic cancer. The phosphoS322 detection antibody might be a valuable tool for predicting pancreatic cancer prognosis. These findings implicate ERK2 as a potential therapeutic target for pancreatic cancer and highlight phosphoZEB1 as a valuable prognostic marker.https://doi.org/10.1186/s12964-025-02182-3Serine 322 phosphorylated ZEB1ERK2Epithelial-mesenchymal transitionSphingosylphosphorylcholinePancreatic cancer |
| spellingShingle | 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 ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression Cell Communication and Signaling Serine 322 phosphorylated ZEB1 ERK2 Epithelial-mesenchymal transition Sphingosylphosphorylcholine Pancreatic cancer |
| title | ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression |
| title_full | ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression |
| title_fullStr | ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression |
| title_full_unstemmed | ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression |
| title_short | ERK2-mediated phosphorylation of ZEB1 at S322 enhances PD-L1 expression and EMT, leading to pancreatic cancer progression |
| title_sort | erk2 mediated phosphorylation of zeb1 at s322 enhances pd l1 expression and emt leading to pancreatic cancer progression |
| topic | Serine 322 phosphorylated ZEB1 ERK2 Epithelial-mesenchymal transition Sphingosylphosphorylcholine Pancreatic cancer |
| url | https://doi.org/10.1186/s12964-025-02182-3 |
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