Journal: Cancer gene therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer.

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: Fig. 1 FOXP1 is upregulated in gemcitabine-resistant (GR) PDAC. A FOXP1 expression levels in PAAD tumor tissues and adjacent normal tissues. FOXP1 is significantly upregulated in tumor tissues. B, C mRNA and protein expression analyses reveal higher FOXP1 expression in PDAC tissue samples than in normal tissues. D Differential gene expression analysis from the GSE71989 dataset reveals high expressions of FOXP1 and CSC markers LEF1 and OCT4 in tumor tissues [21]. E Kaplan–Meier OS and DSS curves for TCGA–pancreatic adenocarcinoma (PAAD) (F) mRNA expression levels of FOXP1 and RRM1, a marker of gemcitabine resistance, in GS and GR tissue samples. G FOXP1 and RRM1 protein expression levels in GR tissues. H, I FOXP1 and RRM1 mRNA and protein expression across GS and GR PDAC cell lines, highlighting higher FOXP1 levels in GR cells, with the highest upregulation in Capan-1. *p < 0.05, **p < 0.01, ***p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [2, 7]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Expressing, Gene Expression, Marker

Journal: Cancer gene therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer.

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: Fig. 2 FOXP1 promotes the proliferation of CSCs in GR PDAC. A Flow cytometry (FACS) analysis of CD44+ CD24+ double-positive cells in gemcitabine-sensitive (GS), gemcitabine-resistant (GR), and FOXP1 knockdown (KD) GR Capan-1 cells. (B) Colony formation assay comparing GS, GR, and GR FOXP1 KD groups. C Western blot analysis of CSC marker genes in monolayer and 3D spheroid cultures of GS and GR Capan-1 cells. D Spheroid formation over 14 days in GS, GR, and GR FOXP1 KD cells. E Quantitative analysis of mRNA levels of CSC markers in GS, GR, and GR FOXP1 KD cells via qRT-PCR. F Western blot of CSC markers in Capan-1 GS and GR cells with and without FOXP1 knockdown. Data are represented as the mean ± S.D. of three independent experiments (n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. **p < 0.01, ***p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [2, 7]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Flow Cytometry, Knockdown, Colony Assay, Western Blot, Marker, Quantitative RT-PCR, Comparison

Journal: Cancer gene therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer.

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: Fig. 4 FOXP1 induces metabolic reprogramming by promoting glycolysis. A GSEA of the TCGA-PAAD cohort shows that high FOXP1 expression is associated with increased glucose metabolism and serum lactate levels. Heatmap analysis of RNA sequencing data from Capan-1 GS and GR cells revealed the upregulation of glycolysis-related genes in GR cells. B Glucose uptake assay showed reduced lactate levels in KD cells compared to GR cells. C mRNA expression analysis of glycolytic genes in GS, GR, and FOXP1 KD GR cells, quantified by qRT-PCR. D Lactate secretion assay showing reduced lactate levels in KD cells compared with GR cells. E mRNA expressions of LDHA and MCT4 in GS, GR, and FOXP1 KD GR cells were quantified by qRT-PCR. F Western blotting analysis of glycolytic enzymes in GS, GR, and KD cells. G OCR and ECAR assays indicated reduced glycolysis in KD cells. Data are represented as the mean ± S.D. of three independent experiments (n = 3). Statistical analyses were conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01.

Article Snippet: siRNA-mediated knockdown was performed as previously described [2, 7]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot, Comparison

Journal: Cancer gene therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer.

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: Fig. 3 FOXP1 enhances EMT and induces proliferation in GR PDAC. A Wound healing assay of GS, GR, and KD Capan-1 cells at 0, 16, and 24 h. Scale bar = 100 μm (B) The number of invaded cells was measured via Transwell Invasion Assay. Scale bar = 100 μm (C) Western blot analysis of epithelial and mesenchymal markers in GS, GR, and FOXP1 KD GR cells. D WST-1 cell viability analysis of GS, GR, and FOXP1 KD GR cells. E Immunofluorescence staining of Ki-67 in GS, GR, and FOXP1 KD GR cells. Scale bar = 50 μm (F) Western blot analysis of activated MAPK protein expression levels in GS, GR, and FOXP1 KD GR cells. G Western blot analysis of expression levels of cell cycle regulators genes in GS, GR, and FOXP1 KD GR cells. Data are represented as the mean ± S.D. of three independent experiments (n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. **p < 0.01, ***p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [2, 7]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Wound Healing Assay, Transwell Invasion Assay, Western Blot, Staining, Expressing, Comparison

Journal: Cancer gene therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer.

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: Fig. 5 FOXP1 chemosensitizes PDAC to gemcitabine by upregulating ABCG2. A Correlation analysis between FOXP1 and ABCG2 expression levels in the TCGA-PAAD dataset via TIMER 2.0. B Western Blot shows reduced ABCG2 expression in FOXP1 KD cells. C WST-1 viability assay of GR cells treated with KO143 at different concentrations. Co-treatment with gemcitabine and KO143 further decreases cell viability. D Sphere formation assay of GS, GR, FOXP1 KD, KO143-treated GR cells, and co-treatment of KO143 and siFOXP1. E FOXP1 binding motif and schematic of the ABCG2 promoter region showing potential FOXP1 binding sites identified via JASPAR. F Primer design for assessing FOXP1 binding to the ABCG2 promoter. G Luciferase reporter assay in HEK293 cells transfected with wild-type (WT) or mutant (MT) ABCG2 promoter constructs. H Chromatin immunoprecipitation (ChIP) assay in HEK293 cells confirms FOXP1 binding to the ABCG2 promoter. Data are represented as the mean ± S.D. of three independent experiments (n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. ns > 0.05, *p < 0.05, **p < 0.01, ***p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [2, 7]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Expressing, Western Blot, Viability Assay, Tube Formation Assay, Binding Assay, Luciferase, Reporter Assay, Transfection, Mutagenesis, Construct, Chromatin Immunoprecipitation, Comparison

Journal: Cancer gene therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer.

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: Fig. 6 Reduced FOXP1 and ABCG2 expressions lead to reduced tumor growth and increased chemosensitivity in mouse models. A Representative image of tumors from BALB/C nude mice injected with GS or GR Capan-1 cells, with GR cells treated with shFOXP1 lentivirus and/or KO143 (Co-Tx). B Quantification of tumor weights after 5 weeks shows significantly smaller tumors in the Co-Tx group. C Western blot analysis of FOXP1 expressions in GS, GR, shFOXP1-treated, KO143-treated, and Co-Tx groups. D Relative mRNA expression levels of stemness markers show significant reductions in the Co-Tx group compared to untreated groups. E, F Immunofluorescence analyses of FOXP1, ABCG2, and stemness marker genes in tumor tissues from each group. Data are represented as the mean ± S.D. of three independent experiments (n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. *p < 0.05, **p < 0.01, ***p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [2, 7]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Injection, Western Blot, Expressing, Marker, Comparison