Journal: Oncogene

Article Title: Partial truncation of the C-terminal domain of PTCH1 in cancer promotes tumourigenesis by non-canonical activation of a GLI-PI3K loop

doi: 10.1038/s41388-026-03698-9

Figure Lengend Snippet: A Top: EREG , AREG , and EGFR mRNA expression in SCR, C9, and C15 cells by qPCR. Expression levels were normalized to GAPDH mRNA and represented as fold change relative to control SCR cells ( n = 3). Bottom: expression of the same genes in pooled RNA from xenografts derived from SCR and C9 cells. B Representative western blot of EGFR, phospho-Akt, phospho-ERK1/2, total Akt, total ERK1/2, and GADPH in SCR, C9, and C15 cells. C Effect of 10 μM Erlotinib or vehicle control on cell proliferation of SCR, C9 and C15 cells over time ( n = 3). D . Effect of 10 μM Erlotinib or vehicle control (DMSO) on the colony formation ability of SCR, C9 and C15 cells upon culture for 10 days. Left: representative images at the end of the experiment. Right: number of colonies per 1,000 plated cells at day 10 ( n = 3). E Effect of 10 μg/ml Cetuximab or a control IgG on the cell number of SCR, C9 and C15 cells after 72 h treatment ( n = 3). F Effect of 30 nM Trametinib or DMSO vehicle on phospho-ERK1/2 levels in SCR, C9 and C15 cells. G Dose dependent reduction of cell proliferation in SCR, C9, and C15 cells treated with Trametinib for 72 h ( n = 3). All quantitative data represent the mean ± SEM of independent biological repeats (indicated n), performed in technical replicates. ns = not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The following antibodies were used: EGFR (Cell Signaling Technology #2256, WB 1:1,000), AKT (Cell Signaling Technology #4691, WB 1:1,000), phospho- AKT (Ser473) (D9E) XP (Cell Signaling Technology #5012, WB 1:1,000), p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology #4695, WB 1:1,000), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology #4376, WB 1:1,000), Gli1 (Cell Signaling Technology #2643, WB 1:1,000), Phospho-PKA substrate (Cell Signaling Technology #9624, WB 1:1,000), GAPDH-HRP (Proteintech #HRP-60004, WB 1:3,000), Vinculin (Santa Cruz #sc-73614, WB 1:10,000), Poly-ADP ribose Polymerase (Cell Signaling Technology #9542, WB 1:2,000), anti VP16 (Santa Cruz #sc7545, WB 1:1,000),HA-Tag (Invitrogen #26183, WB 1:10,000), goat anti-rabbit IgG HRP (BioRad #172-1019, WB 1:3,000), goat anti-mouse IgG (H L)-HRP conjugate (Bethyl #A90-116P, WB 1:3,000).

Techniques: Expressing, Control, Derivative Assay, Western Blot

Journal: Journal of Translational Medicine

Article Title: Integrin αvβ3-dependent pathogenic effect and therapeutic effects of DL-N2 combined with EGFR inhibitors in pancreatic adenocarcinoma

doi: 10.1186/s12967-026-07865-0

Figure Lengend Snippet: Pathway enrichment analysis of ITGAV-associated genes reveals EMT, ECM remodeling, and EGFR-linked signaling. Bar plot shows the top 10 enriched biological pathways associated with ITGAV expression in TCGA-PAAD. Significance is represented as -log(p-value) on the x-axis. ITGAV-associated genes are enriched in pathways related to epithelial-to-mesenchymal transition (EMT), extracellular matrix (ECM) remodeling, and TGF-β-mediated fibroblast migration, processes central to tumor invasion and fibrosis. Several pathways, including tumor–stroma interactions, stellate cell activation, and cytoskeleton remodeling, overlap with EGFR signaling cascades, suggesting cooperative roles of ITGAV and EGFR in promoting pancreatic cancer progression

Article Snippet: Membranes were blocked with 5% skim milk/TBST (TBS with 0.1% Tween-20) and incubated overnight at 4 °C with the following primary antibodies: p-EGFR (1:1000; Cell Signaling #2236), EGFR (1:2000; GeneTex #GTX628887), PD-L1 (1:1000; GeneTex #GTX104763), p-STAT3-Tyr705 (1:1000; Cell Signaling #9145), STAT3 (1:1000; BD Biosciences #610190), p-ERK1/2 (1:1000; Cell Signaling #4377), ERK1/2 (1:2000; Cell Signaling #9102), and GAPDH (1:20,000; Proteintech #60004-1).

Techniques: Expressing, Migration, Activation Assay

Journal: Journal of Translational Medicine

Article Title: Integrin αvβ3-dependent pathogenic effect and therapeutic effects of DL-N2 combined with EGFR inhibitors in pancreatic adenocarcinoma

doi: 10.1186/s12967-026-07865-0

Figure Lengend Snippet: Protein–protein interaction network and correlation of ITGAV and ITGB3 with EGFR expression.( A ) Protein–protein interaction (PPI) network of ITGAV and ITGB3 and their associated genes, generated using the STRING database. The network illustrates key predicted interactions involved in cell adhesion, migration, and epithelial–mesenchymal transition (EMT). Table listing interaction scores between ITGAV/ITGB3 and selected partner genes, highlighting strong associations with EGFR (interaction score > 0.6 for both). ( B–C ) Correlation analysis using GEPIA shows a significant positive correlation between EGFR expression and ITGAV ( B ) as well as EGFR and ITGB3 ( C ) in PAAD samples. Spearman correlation coefficients and p-values are indicated, supporting a functional association between integrin αvβ3 subunits and EGFR signaling in pancreatic cancer

Article Snippet: Membranes were blocked with 5% skim milk/TBST (TBS with 0.1% Tween-20) and incubated overnight at 4 °C with the following primary antibodies: p-EGFR (1:1000; Cell Signaling #2236), EGFR (1:2000; GeneTex #GTX628887), PD-L1 (1:1000; GeneTex #GTX104763), p-STAT3-Tyr705 (1:1000; Cell Signaling #9145), STAT3 (1:1000; BD Biosciences #610190), p-ERK1/2 (1:1000; Cell Signaling #4377), ERK1/2 (1:2000; Cell Signaling #9102), and GAPDH (1:20,000; Proteintech #60004-1).

Techniques: Expressing, Generated, Migration, Functional Assay

Journal: Journal of Translational Medicine

Article Title: Integrin αvβ3-dependent pathogenic effect and therapeutic effects of DL-N2 combined with EGFR inhibitors in pancreatic adenocarcinoma

doi: 10.1186/s12967-026-07865-0

Figure Lengend Snippet: DL-N2 regulates gene expression and enhances gefitinib-induced inhibition in pancreatic cancer cells. ( A–D ) BxPC-3 and ( E - H ) AsPC-1 cells treated with DL-N2 (10⁻⁹–10⁻⁷ M), gefitinib (10 µM), or their combinations on pancreatic cancer cells. Quantitative real-time PCR analysis of EGFR ( A and E ), PD-L1 ( B and F ), CCND1 ( C and G ), and PCNA ( D and H ) expression. DL-N2 decreased the expression of proliferation- and immune-related genes, with maximal inhibition observed in combination with gefitinib. Data are presented as mean ± SD of four independent experiments ( N = 4). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. gefitinib alone

Article Snippet: Membranes were blocked with 5% skim milk/TBST (TBS with 0.1% Tween-20) and incubated overnight at 4 °C with the following primary antibodies: p-EGFR (1:1000; Cell Signaling #2236), EGFR (1:2000; GeneTex #GTX628887), PD-L1 (1:1000; GeneTex #GTX104763), p-STAT3-Tyr705 (1:1000; Cell Signaling #9145), STAT3 (1:1000; BD Biosciences #610190), p-ERK1/2 (1:1000; Cell Signaling #4377), ERK1/2 (1:2000; Cell Signaling #9102), and GAPDH (1:20,000; Proteintech #60004-1).

Techniques: Gene Expression, Inhibition, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: Journal of Translational Medicine

Article Title: Integrin αvβ3-dependent pathogenic effect and therapeutic effects of DL-N2 combined with EGFR inhibitors in pancreatic adenocarcinoma

doi: 10.1186/s12967-026-07865-0

Figure Lengend Snippet: DL-N2 and gefitinib cooperatively suppress EGFR activation and downstream signaling in pancreatic cancer cells. ( A - B ) Western blot and densitometric analyses show the effects of DL-N2 (10⁻⁹–10⁻⁷ M) and gefitinib (10 µM) on EGFR-related signaling in BxPC-3 cellsTreatment with DL-N2 alone modestly reduced pEGFR and PD-L1 expression, whereas co-treatment with gefitinib further decreased pEGFR and PD-L1 levels compared with either agent alone. Total EGFR remained largely unchanged. Analysis of STAT3 signaling shows that DL-N2 alone transiently increased pSTAT3-Tyr705, while combination treatment with gefitinib suppressed pERK1/2 activation without markedly altering total STAT3 expression. Data are presented as mean ± SD of four independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 versus untreated control

Article Snippet: Membranes were blocked with 5% skim milk/TBST (TBS with 0.1% Tween-20) and incubated overnight at 4 °C with the following primary antibodies: p-EGFR (1:1000; Cell Signaling #2236), EGFR (1:2000; GeneTex #GTX628887), PD-L1 (1:1000; GeneTex #GTX104763), p-STAT3-Tyr705 (1:1000; Cell Signaling #9145), STAT3 (1:1000; BD Biosciences #610190), p-ERK1/2 (1:1000; Cell Signaling #4377), ERK1/2 (1:2000; Cell Signaling #9102), and GAPDH (1:20,000; Proteintech #60004-1).

Techniques: Activation Assay, Western Blot, Expressing, Control

Journal: Journal of Translational Medicine

Article Title: Integrin αvβ3-dependent pathogenic effect and therapeutic effects of DL-N2 combined with EGFR inhibitors in pancreatic adenocarcinoma

doi: 10.1186/s12967-026-07865-0

Figure Lengend Snippet: Pathway enrichment analysis of ITGB3-associated genes highlights EMT, fibroblast activation, and EGFR pathway crosstalk. Bar plot shows the top 10 enriched biological pathways associated with ITGB3 expression in TCGA-PAAD. Significance is represented as -log(p-value) on the x-axis. ITGB3-associated genes are enriched in EMT, ECM remodeling, and TGF-β-driven fibroblast migration pathways that contribute to desmoplasia and metastasis. Enriched pathways related to stellate cell activation, cytoskeleton reorganization, and tumor–stroma signaling also align with EGFR-linked oncogenic processes, indicating integrin–EGFR pathway synergy within the pancreatic tumor microenvironment

Article Snippet: Membranes were blocked with 5% skim milk/TBST (TBS with 0.1% Tween-20) and incubated overnight at 4 °C with the following primary antibodies: p-EGFR (1:1000; Cell Signaling #2236), EGFR (1:2000; GeneTex #GTX628887), PD-L1 (1:1000; GeneTex #GTX104763), p-STAT3-Tyr705 (1:1000; Cell Signaling #9145), STAT3 (1:1000; BD Biosciences #610190), p-ERK1/2 (1:1000; Cell Signaling #4377), ERK1/2 (1:2000; Cell Signaling #9102), and GAPDH (1:20,000; Proteintech #60004-1).

Techniques: Activation Assay, Expressing, Migration

Journal: Journal of Translational Medicine

Article Title: Integrin αvβ3-dependent pathogenic effect and therapeutic effects of DL-N2 combined with EGFR inhibitors in pancreatic adenocarcinoma

doi: 10.1186/s12967-026-07865-0

Figure Lengend Snippet: Conclusion summarizing the role of integrin αvβ3 and therapeutic potential of DL-N2 in pancreatic cancer. Schematic diagram depicting how integrin αvβ3 cooperates with EGFR and downstream signaling cascades to promote tumor progression. Integrin αvβ3 facilitates EGF-mediated activation of the FAK–Src–Ras–ERK and PI3K–AKT pathways, driving proliferation, survival, migration, and immune evasion through EMT regulators (SNAI1, ZEB1), transcriptional control of PD-L1, and extracellular matrix remodeling factors (MMP2, MMP9, FN1, CD44). Crosstalk with TGF-β/SMAD2/3 further may may enhances invasion and drug resistance. DL-N2, a tetrac derivative, disrupts integrin αvβ3 signaling, reduces ERK activation, restores chemosensitivity, and suppresses PD-L1 expression. In combination with gefitinib or as DL-N2–doxorubicin (DL-N2-Dox), the compound may may enhances. DNA damage responses, increases ROS, and induces apoptosis while attenuating EMT, immune evasion, and tumor–stroma interactions. This integrative model highlights integrin αvβ3 as a therapeutic target and DL-N2 as a promising strategy for overcoming drug resistance in pancreatic cancer

Article Snippet: Membranes were blocked with 5% skim milk/TBST (TBS with 0.1% Tween-20) and incubated overnight at 4 °C with the following primary antibodies: p-EGFR (1:1000; Cell Signaling #2236), EGFR (1:2000; GeneTex #GTX628887), PD-L1 (1:1000; GeneTex #GTX104763), p-STAT3-Tyr705 (1:1000; Cell Signaling #9145), STAT3 (1:1000; BD Biosciences #610190), p-ERK1/2 (1:1000; Cell Signaling #4377), ERK1/2 (1:2000; Cell Signaling #9102), and GAPDH (1:20,000; Proteintech #60004-1).

Techniques: Activation Assay, Migration, Control, Expressing