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<p>Local invasion is considered a premonitor of tumor metastasis which cause curative difficulties and undesired prognosis in patients with Pancreatic ductal adenocarcinoma (PDAC). The importance of mRNA N6-methyladenosine (m<sup>6</sup>A) modification during tumor invasion is controversial as it plays distinct roles which mainly attributed to different m<sup>6</sup>A reader proteins exert function. In current study, the level of m<sup>6</sup>A expression in PDAC was analyzed by IHC and ELISA, all m<sup>6</sup>A-regulated genes in PDAC detected by qPCR. The downstream gene <i>EMP1</i> was screened by analyzing <i>IGF2BP3</i> knockdown RNA-seq, <i>IGF2BP3</i>-RIP, MeRIP-seq, and PDAC-survival related genes. And m<sup>6</sup>A modification sites of <i>EMP1</i> RNA was verified by MeRIP-qPCR, RIP-qPCR, and dual luciferase assays. <i>EMP1</i>-binding protein <i>VASP</i> was identified by mass spectrometry. Cell migration and invasive activity were detected using cytoskeletal staining, scratch assay, transwell assay, subcutaneous tumor and lung metastasis models. Finally, prognosis and immune microenvironment was analyzed in PDAC by IHC and multiple immunofluorescence staining. This work shows that m<sup>6</sup>A reader <i>IGF2BP3</i> is remarkably upregulated in local invasion PDAC and indicates worse prognosis of patients. Mechanistically, <i>IGF2BP3</i> recognized m<sup>6</sup>A-modified <i>EMP1</i> mRNAs to prolong stability of them, which inhibits the hindrance of <i>SMAD7</i> to <i>SMAD3/4</i> phosphorylation by promoting the binding of <i>VASP</i> and <i>SMAD7</i>. Finally, a tight correlation of the local invasion/<i>IGF2BP3/EMP1</i> and infiltration of immune cells in the tumor microenvironment is evidenced in clinical PDAC. In conclusions, <i>IGF2BP3</i> functions as an invasion driver that induces PDAC development via the <i>EMP1/TGF-β</i> axis. And <i>IGF2BP3/EMP1</i> axis may be involved in regulating microenvironmental remodeling in pancreatic cancer.</p>

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IGF2BP3 regulates EMP1 stability in an m6A-dependent manner and activates the TGF-β pathway to promote pancreatic cancer invasion

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年11月24日

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  • Long Liu,
  • Yu Zhang,
  • Yuxi Huang,
  • Xiaohong Zhao,
  • Hongxing Fu,
  • Lin Chen,
  • Qi Wang,
  • Xingyu Liu,
  • Xiang Zheng,
  • Hengqing Zhu,
  • Zheping Fang,
  • Zhenzhen Gao,
  • Sheng Yan

摘要

Local invasion is considered a premonitor of tumor metastasis which cause curative difficulties and undesired prognosis in patients with Pancreatic ductal adenocarcinoma (PDAC). The importance of mRNA N6-methyladenosine (m6A) modification during tumor invasion is controversial as it plays distinct roles which mainly attributed to different m6A reader proteins exert function. In current study, the level of m6A expression in PDAC was analyzed by IHC and ELISA, all m6A-regulated genes in PDAC detected by qPCR. The downstream gene EMP1 was screened by analyzing IGF2BP3 knockdown RNA-seq, IGF2BP3-RIP, MeRIP-seq, and PDAC-survival related genes. And m6A modification sites of EMP1 RNA was verified by MeRIP-qPCR, RIP-qPCR, and dual luciferase assays. EMP1-binding protein VASP was identified by mass spectrometry. Cell migration and invasive activity were detected using cytoskeletal staining, scratch assay, transwell assay, subcutaneous tumor and lung metastasis models. Finally, prognosis and immune microenvironment was analyzed in PDAC by IHC and multiple immunofluorescence staining. This work shows that m6A reader IGF2BP3 is remarkably upregulated in local invasion PDAC and indicates worse prognosis of patients. Mechanistically, IGF2BP3 recognized m6A-modified EMP1 mRNAs to prolong stability of them, which inhibits the hindrance of SMAD7 to SMAD3/4 phosphorylation by promoting the binding of VASP and SMAD7. Finally, a tight correlation of the local invasion/IGF2BP3/EMP1 and infiltration of immune cells in the tumor microenvironment is evidenced in clinical PDAC. In conclusions, IGF2BP3 functions as an invasion driver that induces PDAC development via the EMP1/TGF-β axis. And IGF2BP3/EMP1 axis may be involved in regulating microenvironmental remodeling in pancreatic cancer.