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Article Open Access

Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation

  • Authors:
    • Wenjing Yu
    • Junqun Liao
    • Jiayan Li
    • Yongquan Wu
  • View Affiliations / Copyright

    Affiliations: The Medical Laboratory of Science, The Thirteenth People's Hospital of Chongqing, Chongqing 400084, P.R. China, The Medical Laboratory of Science, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400084, P.R. China, Zhongyuan Huiji Biotechnology Co., Ltd. (Zybio Inc.), Chongqing 400084, P.R. China
    Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 602
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    Published online on: October 21, 2025
       https://doi.org/10.3892/ol.2025.15348
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Abstract

The stimulator of interferon genes (STING) signaling pathway plays an important role in tumor progression, particularly in immune regulation. However, the functions of STING‑related genes in immunologically ‘cold’ tumors, such as pancreatic ductal adenocarcinoma (PAAD), are yet to be elucidated. The present study aims to investigate the involvement of STING signaling pathway‑associated genes in PAAD progression and immune infiltration. Bioinformatics analysis of PAAD transcriptomic data from public cohorts (TCGA and GEO) revealed that the majority of STING‑related genes were highly expressed in PAAD. Based on the expression levels of these genes, patients with PAAD from these cohorts were stratified into high‑risk and low‑risk groups. These STING‑related genes exerted differential impacts on tumor mutational burden, immune infiltration and response to immunotherapy. Utilizing these genes, a prognostic model was constructed (hazard ratio, 2.639; P<0.001) and externally validated. Model analysis indicated that interferon‑induced protein with tetratricopeptide repeats 2 (IFIT2) and zinc finger DHHC‑type containing 1 (ZDHHC1) may possess notable biological functions in PAAD. Subsequent functional experiments involving knockout of IFIT2 and overexpression of ZDHHC1in PAAD cell lines demonstrated that these genes significantly regulated cell proliferation (assessed by CCK‑8 assay) and apoptosis (assessed by TUNEL assay) (P<0.001). In summary, the STING signaling pathway is critically involved in PAAD pathogenesis, and the representative genes IFIT2 and ZDHHC1 exhibit distinct functional roles within this context.
View Figures

Figure 1

Role of STING expression in PAAD. (A)
Heatmap comparing STING and immune-related gene expression profiles
in PAAD tumor microenvironment vs. adjacent normal tissue. (B) Box
plot demonstrating significantly elevated STING mRNA expression in
PAAD tumors (T) vs. normal pancreatic tissues (N) (*P<0.05)
(TCGA-PAAD (T) + Genotype-Tissue Expression; normal controls;
http://gtexportal.org). (C) Kaplan-Meier curves
comparing overall survival between PAAD patients with low vs. high
STING expression (log-rank test). (D) Violin plots showing STING
expression distribution across AJCC stages (I–IV) of PAAD. (E)
Oncoprint of STING genomic alterations (mutations, copy number
changes) in PAAD. (F) Human Protein Atlas representative
immunohistochemistry images of STING protein expression (PAAD
tissue; proteinatlas.org). Scale bar, 100 µm. STING, stimulator of
interferon genes; PAAD, pancreatic ductal adenocarcinoma; PAAD,
pancreatic adenocarcinoma; IHC, immunohistochemistry; NK, natural
killer; HR, hazard ratio; T, tumor; N, normal; TPM, transcripts per
million.

Figure 2

Molecular characterization of STING
signaling pathway-related genes in PAAD. (A) Protein-protein
interaction (PPI) network of core STING signaling components
identified through STRING database analysis (minimum interaction
score: 0.4). (B) Expanded PPI network showing secondary interactors
of STING pathway genes (minimum interaction score: 0.7). (C)
Consensus matrix heatmap classifying patients with PAAD into two
molecular subtypes (C1 and C2) based on STING-related gene
expression patterns. (D) Heatmap comparing expression profiles of
STING-related genes across PAAD subtypes (C1 vs. C2) with
expression gradient key (red, high expression; blue, low
expression). (E) Kaplan-Meier survival analysis showing
significantly reduced overall survival in the STING-enriched
subtype (C1) compared with the STING-depleted subtype (C2). STING,
stimulator of interferon genes; PAAD, pancreatic ductal
adenocarcinoma; CGAS, cyclic GMP-AMP synthase; TBK1, TANK-binding
kinase 1; MAVS, mitochondrial antiviral-signaling protein; TRIM56,
tripartite motif-containing 56; TREX1, three prime repair
exonuclease 1; TRIM32, tripartite motif-containing 32; IFI16,
interferon γ inducible protein 16; IFIT1, interferon-induced
protein with tetratricopeptide repeats 1; DDX41, DEAD-box helicase
41; NLRP4, NLR family pyrin domain containing 4; DTX4, deltex E3
ubiquitin ligase 4; ZDHHC1, zinc finger DHHC-type containing 1;
PRKDC, protein kinase DNA-activated catalytic subunit; XRCC, X-ray
repair cross complementing; ELF4, E74-like ETS transcription factor
4; NLRC3, NLR family CARD domain containing 3; ARF1, ARF GTPase 1;
DDX56, RNA sensor RIG-I; IKBKE, inhibitor of nuclear factor κB
kinase subunit ε; IRF3, interferon regulatory factor 3; RNF5, ring
finger protein 5.

Figure 3

Gene characteristics of different
PAAD subtypes. (A) Heatmap of upregulated genes in STING1-enriched
subtype (C1) vs. STING1-depleted subtype (C2). (B) Heatmap of
downregulated genes in STING1-enriched subtype (C1) vs.
STING1-depleted subtype (C2). (red, high expression; blue, low
expression). (C) GO enrichment analysis showing top biological
processes in STING1-enriched subtype. (D) KEGG pathway enrichment
analysis showing top signaling pathways in STING1-enriched subtype.
(E) Mutational landscape of STING1-enriched subtype (C1) showing
frequent mutations in KRAS, TP53 and CDKN2A. (F) Mutational
landscape of STING1-depleted subtype (C2) showing distinct mutation
profile with SMAD4 alterations. PAAD, pancreatic ductal
adenocarcinoma; STING, stimulator of interferon genes; FC,
fold-change; fdr, false discovery rate; TMB, tumor mutational
burden; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and
Genomes.

Figure 4

Immune microenvironment profiling of
PAAD molecular subtypes. (A) ESTIMATE analysis showing
significantly higher immune scores in STING1-high subtype (C1) vs.
STING1-low subtype (C2). (B) Significantly higher stromal scores in
C1 vs. C2. (C) Significantly lower tumor purity in C1 vs. C2. (D)
CIBERSORT analysis revealing differential immune cell infiltration
patterns between subtypes (key immune subsets labeled)
(***P<0.001). (E) Positive correlation between STING1 expression
and γδ T cell infiltration density (Pearson r=0.68; *P<0.05) (F)
Upregulated HLA molecule expression in C1 vs. C2 (**P<0.05 and
***P<0.01). (G) Elevated immune checkpoint molecule expression
in C1 vs. C2. PAAD, pancreatic ductal adenocarcinoma; STING,
stimulator of interferon genes; HLA, human leukocyte antigen; NK,
natural killer.

Figure 5

Development and validation of a
STING1 signaling-related prognostic signature for PAAD. (A)
Univariate Cox regression analysis of STING1 pathway-related genes
in PAAD. Genes with significant prognostic value (P<0.05) are
included. (B) LASSO regression 10-fold cross-validation curve
showing optimal λ selection (minimum criteria). (C) LASSO
coefficient profiles of STING1-related genes across log(λ)
sequence. (D) Kaplan-Meier survival analysis for the prognostic
signature in TCGA-PAAD cohort (high-risk vs. low-risk groups;
cutoff: median risk score). (E) Validation of prognostic signature
in GSE224564 cohort. (F) Risk score distribution plot with
corresponding survival status and expression heatmap for 6
signature genes in TCGA-PAAD. (G) Validation in GSE224564 cohort
showing identical configuration to (F). STING, stimulator of
interferon genes; PAAD, pancreatic ductal adenocarcinoma; LASSO,
least absolute shrinkage and selection operator; TCGA, The Cancer
Genome Atlas; MAVS, mitochondrial antiviral-signaling protein;
IFI16, interferon γ inducible protein 16; IFIT2, interferon-induced
protein with tetratricopeptide repeats 2; DTX4, deltex E3 ubiquitin
ligase 4; ZDHHC1, zinc finger DHHC-type containing 1; PRKDC,
protein kinase DNA-activated catalytic subunit; STAT6, signal
transducer and activator of transcription 6; XRCC5, X-ray repair
cross complementing: ELF4, E74-like ETS transcription factor 4;
DDX58, RNA sensor RIG-I.

Figure 6

Independent prognostic value and
immunotherapy relevance of the STING-related risk signature in
PAAD. (A) Univariate Cox regression analysis of clinical parameters
and risk score. (B) Multivariate Cox regression confirming the risk
score as an independent prognostic factor after adjusting for
covariates in (A). (C) Correlation analysis of risk score and
activated dendritic cell abundance. ‘Dendritic cells activated’
represents proportion scores derived from the xCell algorithm (0–1
scale), quantifying relative abundance in the tumor
microenvironment. Correlation was assessed using Spearman's rank
correlation analysis (Spearman's ρ=0.28; P=0.015). (D) Higher risk
scores in immunotherapy non-responders vs. responders (Mann-Whitney
U test). Single-cell RNA sequencing (GSE141017) visualization of
STING1 signature gene expression: (E) t-SNE plot showing cell-type
distribution. (F) Feature plots of signature gene expression across
the PAAD tumor microenvironment. STING, stimulator of interferon
genes; PAAD, pancreatic ductal adenocarcinoma; T, tumor; N, node;
CD8T, T-cell surface glycoprotein CD8; ZDHHC1, zinc finger
DHHC-type containing 1; IFIT2, interferon-induced protein with
tetratricopeptide repeats 2; IFI16, interferon γ inducible protein
16; DTX4, deltex E3 ubiquitin ligase 4; MAVS, mitochondrial
antiviral-signaling protein; PRKDC, protein kinase DNA-activated
catalytic subunit; PAAD, pancreatic adenocarcinoma.

Figure 7

Functional roles of IFIT2 and ZDHHC1
in PAAD oncobiology. (A) Human Protein Atlas representative
immunohistochemical staining of six prognostic signature genes in
PAAD tissues (proteinatlas.org). Scale bar, 100 µm. (B) Western
blot and semi-quantification confirming successful ZDHHC1
overexpression in Panc02 cells (C) Western blot and
semi-quantification confirming IFIT2 knockout efficiency in Panc02
cells (****P<0.0001). (D) CCK-8 assay: ZDHHC1 overexpression
enhances Panc02 proliferation (***P<0.001 and ****P<0.0001 vs
OE-NC). (E) CCK-8 assay: IFIT2 knockout enhances Panc02
proliferation (**P<0.01 and ***P<0.001 vs. sh-NC). (F) TUNEL
assay (scale bar, 50 µm) demonstrating reduced apoptosis in
ZDHHC1-overexpressing Panc02 cells. (G) TUNEL assay (scale bar 50
µm) showing decreased apoptosis in IFIT2-knockout Panc02 cells
(**P<0.01). IFIT2, interferon-induced protein with
tetratricopeptide repeats 2; ZDHHC1, zinc finger DHHC-type
containing 1; PAAD, pancreatic ductal adenocarcinoma; OE,
overexpression; CCK-8, Cell Counting Kit-8; DTX4, deltex E3
ubiquitin ligase 4; IFI16, interferon γ inducible protein 16; MAVS,
mitochondrial antiviral-signaling protein; PRKDC, protein kinase
DNA-activated catalytic subunit; sh-NC, scrambled negative control
shRNA; ns, not significant.
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Copy and paste a formatted citation
Spandidos Publications style
Yu W, Liao J, Li J and Wu Y: Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation. Oncol Lett 30: 602, 2025.
APA
Yu, W., Liao, J., Li, J., & Wu, Y. (2025). Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation. Oncology Letters, 30, 602. https://doi.org/10.3892/ol.2025.15348
MLA
Yu, W., Liao, J., Li, J., Wu, Y."Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation". Oncology Letters 30.6 (2025): 602.
Chicago
Yu, W., Liao, J., Li, J., Wu, Y."Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation". Oncology Letters 30, no. 6 (2025): 602. https://doi.org/10.3892/ol.2025.15348
Copy and paste a formatted citation
x
Spandidos Publications style
Yu W, Liao J, Li J and Wu Y: Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation. Oncol Lett 30: 602, 2025.
APA
Yu, W., Liao, J., Li, J., & Wu, Y. (2025). Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation. Oncology Letters, 30, 602. https://doi.org/10.3892/ol.2025.15348
MLA
Yu, W., Liao, J., Li, J., Wu, Y."Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation". Oncology Letters 30.6 (2025): 602.
Chicago
Yu, W., Liao, J., Li, J., Wu, Y."Stimulator of interferon genes signaling network‑driven prognostic signature for pancreatic cancer: Hub gene discovery with multimodal validation". Oncology Letters 30, no. 6 (2025): 602. https://doi.org/10.3892/ol.2025.15348
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