Abstract
Objective
Chronic pancreatitis (CP) creates an inflammatory pancreatic microenvironment and is an established risk factor for pancreatic cancer (PC). However, immune-inflammatory links between CP and PC remain unclear. Isorhamnetin (ISO), a natural flavonoid with anti-inflammatory and antitumor activities, may influence pancreatic inflammatory signaling. This study investigated ISO-associated molecular networks shared by CP and PC, examined the relationships between prioritized targets and transcriptome-estimated immune-cell infiltration, and evaluated the effects of ISO in pancreatic cancer cells.
Methods
CP- and PC-associated targets were collected from disease databases, and ISO targets were obtained from drug-target prediction platforms. Disease- and ISO-related targets were analyzed using protein–protein interaction networks, functional enrichment, and hub-gene screening. The expression, prognostic relevance, diagnostic performance, and immune infiltration associations of hub genes were evaluated using TCGA-PAAD, GTEx, Kaplan–Meier Plotter, ROC analysis, and CIBERSORTx. Molecular docking and 100 ns molecular dynamics simulations assessed ISO–target binding stability. Two-sample Mendelian randomization (MR) explored potential causal associations between genetically proxied circulating levels of the six hub proteins and CP or PC risk. The effects of ISO on proliferation, wound closure, apoptosis, and target protein expression were examined in PANC-1 and BXPC-3 cells.
Results
372 overlapping CP- and PC-associated targets and 48 ISO-related targets were identified. EGFR, AKT1, MMP9, BCL2, TNF, and IL6 were screened as hub genes. Functional enrichment indicated targets were involved in inflammatory responses, cytokine-related signaling, apoptosis regulation, epithelial proliferation, pancreatic cancer-related pathways, and PI3K-Akt signaling. Hub-gene expression was correlated with CIBERSORTx-estimated immune-cell fractions, including activated natural killer cells and several T-cell subsets. Molecular docking and molecular dynamics simulations provided preliminary support for ISO–target interactions and complex stability. Across the 12 prespecified protein–outcome combinations, MMP9–CP was the only nominal IVW association.
In vitro
, ISO reduced the viability and wound closure of PANC-1 and BXPC-3 cells, promoted apoptosis, and decreased EGFR, AKT1, BCL2, and TNF-α protein expression in a concentration-dependent manner.
Conclusion
ISO may influence CP- and PC-associated molecular networks through multitarget regulation involving EGFR, AKT1, MMP9, BCL2, TNF, and IL6. These findings provide hypothesis-generating computational, genetic, and cellular evidence supporting investigation of ISO in pancreatic inflammation-associated tumor biology. However, immune regulatory effects, MMP9/IL6 protein modulation, and EGFR/AKT pathway activation require further experimental validation.