Journal: Discover Oncology

Article Title: Circ_0041150 inhibits proliferation of pancreatic adenocarcinoma cells by regulating triglyceride accumulation via the miR-1178-3p/AADAC axis

doi: 10.1007/s12672-026-04577-1

Figure Lengend Snippet: Analysis of circ_0041150 expression and its impact on PAAD cell proliferation. A The chromosomal location and structure of circ_0041150 were depicted. B The expression patterns of circ_0041150 in HPDE6-C7 and PAAD cell lines (Capan-2, BxPC-3, AsPC-1, PANC-1) were examined by qRT-PCR ( n = 4). C , D The stability of circ_0041150, compared to its host gene RPH3AL mRNA, was analyzed by qRT-PCR after RNase R digestion ( n = 4). E , F Its subcellular localization was assessed via qRT-PCR ( n = 4). G BxPC-3 cells were transfected with sh-circ_0041150#1 or sh-circ_0041150#2, and circ_0041150 expression was detected by qRT-PCR ( n = 4). H , I Cell proliferation in BxPC-3 cells with circ_0041150 knockdown was evaluated by CCK-8 and EdU incorporation assays ( n = 4). J PANC-1 cells were transfected with p-circ_0041150, and circ_0041150 expression was detected by qRT-PCR ( n = 4). K , L Cell proliferation in PANC-1 cells overexpressing circ_0041150 was assessed by CCK-8 and EdU incorporation assays ( n = 4). Statistical significance was assessed using two-tailed unpaired Student’s t-test or two-way ANOVA. * P < 0.05 and ** P < 0.01 vs. control

Article Snippet: The human pancreatic duct epithelial cell line (HPDE6-C7) and PAAD cell lines (Capan-2, BxPC-3, AsPC-1, PANC-1) were obtained from ATCC.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Knockdown, CCK-8 Assay, Two Tailed Test, Control

Journal: Discover Oncology

Article Title: Circ_0041150 inhibits proliferation of pancreatic adenocarcinoma cells by regulating triglyceride accumulation via the miR-1178-3p/AADAC axis

doi: 10.1007/s12672-026-04577-1

Figure Lengend Snippet: Effects of circ_0041150 on TG accumulation of PAAD cells. A Flutamide hydrolase activity was assayed in BxPC-3 cells following circ_0041150 knockdown ( n = 4). B , C Lipid deposition, induced by FFA, was visualized in BxPC-3 cells with knocked-down circ_0041150 via BODIPY 493/503 staining using fluorescence microscopy (40×; scale bar = 20 μm) ( n = 4). D Triglyceride (TG) content was quantified in BxPC-3 cells post-knockdown. E Flutamide hydrolase activity was assayed in PANC-1 cells following circ_0041150 overexpression ( n = 4). F , G FFA-induced lipid deposition was visualized in PANC-1 cells with overexpressed circ_0041150 via BODIPY 493/503 staining using fluorescence microscopy (40×; scale bar = 20 μm) ( n = 4). H TG content was quantified in PANC-1 cells post-overexpression ( n = 4). Statistical significance was assessed using two-tailed unpaired Student’s t-test or two-way ANOVA. * P < 0.05 and ** P < 0.01 vs. control

Article Snippet: The human pancreatic duct epithelial cell line (HPDE6-C7) and PAAD cell lines (Capan-2, BxPC-3, AsPC-1, PANC-1) were obtained from ATCC.

Techniques: Activity Assay, Knockdown, Staining, Fluorescence, Microscopy, Over Expression, Two Tailed Test, Control

Journal: Discover Oncology

Article Title: Circ_0041150 inhibits proliferation of pancreatic adenocarcinoma cells by regulating triglyceride accumulation via the miR-1178-3p/AADAC axis

doi: 10.1007/s12672-026-04577-1

Figure Lengend Snippet: Circ_0041150 targets and regulates miR-1178-3p in PAAD cells. A , B qRT-PCR analysis compared the expression levels of miR-1178-3p, miR-1184, miR-188-3p, and miR-766 in BxPC-3 and PANC-1 cells transfected with sh-circ_0041150#1 or p-circ_0041150 to controls ( n = 4). C Bioinformatics analysis predicted binding site sharing between circ_0041150 and miR-1178-3p, with wild-type and mutated sequences highlighted, with mutation site in red. D , E FISH analysis confirmed co-localization of circ_0041150 (red) and miR-1178-3p (green) in BxPC-3 (D) and PANC-1 (E) cells, with nuclei counterstained with DAPI (blue) (20×; scale bar = 10 μm). F , G Luciferase reporter assays demonstrated altered activity in BxPC-3 (F) and PANC-1 (G) cells transfected with wild-type (WT) or mutant (MUT) circ_0041150 reporters alongside miR-1178-3p mimic or control mimic ( n = 4). H , I RNA pull-down assays followed by qPCR detected circ_0041150 enrichment in BxPC-3 (H) and PANC-1 (I) cell lysates using biotin-labeled miR-1178-3p or control probes ( n = 4). Controls include: (1) bio-NC probe (negative control, a non-specific biotin-labeled miRNA probe with no homology to miR-1178-3p); (2) bio-miR-1178-3p probe (MUT) (mutant control, biotin-labeled miR-1178-3p probe with a mutated binding site for AADAC mRNA, used to verify the specificity of the interaction between miR-1178-3p and AADAC mRNA). J , K AGO2-RIP followed by qPCR assessed circ_0041150 enrichment in BxPC-3 (J) and PANC-1 (K) cells transfected with miR-1178-3p mimic or control mimic ( n = 4). Controls include: (1) Antibody control: Normal rabbit IgG (the IgG subgroup under each transfection condition), which is paired with anti-AGO2 antibody in every transfection group, serving to exclude non-specific immunoprecipitation signals irrelevant to AGO2; (2) Transfection control: control mimic (a non-specific miRNA mimic with no homology to miR-1178-3p, corresponding to the control mimic group in the assay), used to eliminate non-specific effects caused by miRNA mimic transfection itself (instead of the specific function of miR-1178-3p). Statistical significance was assessed using two-tailed unpaired Student’s t-test or two-way ANOVA. * P < 0.05 and ** P < 0.01 vs. control

Article Snippet: The human pancreatic duct epithelial cell line (HPDE6-C7) and PAAD cell lines (Capan-2, BxPC-3, AsPC-1, PANC-1) were obtained from ATCC.

Techniques: Quantitative RT-PCR, Expressing, Transfection, Binding Assay, Mutagenesis, Luciferase, Activity Assay, Control, Labeling, Negative Control, Immunoprecipitation, Two Tailed Test

Journal: Discover Oncology

Article Title: Circ_0041150 inhibits proliferation of pancreatic adenocarcinoma cells by regulating triglyceride accumulation via the miR-1178-3p/AADAC axis

doi: 10.1007/s12672-026-04577-1

Figure Lengend Snippet: AADAC is a direct target of miR-1178-3p in PAAD cells. A Potential binding site overlap between the 3’ UTR of AADAC and miR-1178-3p was revealed by bioinformatics analysis. B , C Luciferase activity was assessed in reporter plasmids containing wild-type and mutant AADAC 3’ UTR sequences co-transfected with miR-1178-3p mimic or negative control in BxPC-3 (B) and PANC-1 (C) cells ( n = 4). D , E AADAC mRNA levels were quantified in pulldown products from cell lysates of BxPC-3 (D) and PANC-1 (E) cells incubated with biotin-labeled miR-1178-3p or control probes ( n = 4). F , G AADAC mRNA levels were detected in BxPC-3 (F) and PANC-1 (G) cells transfected with miR-1178-3p mimic or control mimic ( n = 4). H , I miR-1178-3p and AADAC mRNA expressions were detected in BxPC-3 cells transfected with miR-1178-3p mimic or control mimic ( n = 4). J Following the relevant transfections, AADAC protein expression levels in BxPC-3 cells were evaluated by Western blot ( n = 3). Lane 1, control mimic; Lane 2, miR-1178-3p mimic. K , L miR-1178-3p and AADAC mRNA expressions were also detected in PANC-1 cells transfected with miR-1178-3p inhibitor or miRNA inhibitor-NC ( n = 4). M Following the relevant transfections, AADAC protein expression levels in PANC-1 cells were evaluated by Western blot ( n = 3). Lane 1, miRNA inhibitor-NC; Lane 2, miR-1178-3p inhibitor. Statistical significance was assessed using two-tailed unpaired Student’s t-test or two-way ANOVA. * P < 0.05 and ** P < 0.01 vs. control

Article Snippet: The human pancreatic duct epithelial cell line (HPDE6-C7) and PAAD cell lines (Capan-2, BxPC-3, AsPC-1, PANC-1) were obtained from ATCC.

Techniques: Binding Assay, Luciferase, Activity Assay, Mutagenesis, Transfection, Negative Control, Incubation, Labeling, Control, Expressing, Western Blot, Two Tailed Test

Journal: Discover Oncology

Article Title: Circ_0041150 inhibits proliferation of pancreatic adenocarcinoma cells by regulating triglyceride accumulation via the miR-1178-3p/AADAC axis

doi: 10.1007/s12672-026-04577-1

Figure Lengend Snippet: A schematic model summarizes the study results. In the cytoplasm, circ_0041150 acts as a sponge for miR-1178-3p, preventing its binding to the 3’ UTR of AADAC mRNA, thus increasing AADAC expression, inhibiting TG accumulation and reducing PAAD cell proliferation. Conversely, without circ_0041150, miR-1178-3p binds to AADAC mRNA, inhibiting its expression, promoting TG accumulation and enhancing PAAD cell proliferation

Article Snippet: The human pancreatic duct epithelial cell line (HPDE6-C7) and PAAD cell lines (Capan-2, BxPC-3, AsPC-1, PANC-1) were obtained from ATCC.

Techniques: Binding Assay, Expressing

Journal: Neoplasia (New York, N.Y.)

Article Title: Interdependence of Gemcitabine Treatment, Transporter Expression, and Resistance in Human Pancreatic Carcinoma Cells 1

doi:

Figure Lengend Snippet: MRP5 and gemcitabine chemosensitivity of HEK and PANC-1 cells. Parental HEK cells (open triangles, upper panel) and MRP5-overexpressing HEK/MRP5 cells (filled triangles, upper panel) or PANC-1 cells (open circles, lower panel) and MRP5-overexpressing PANC-1/MRP5 cells (filled circles, lower panel) were subjected to gemcitabine (HEK: 0–100 nM for 4 days; PANC-1: 0–10 µM for 6 days) before cytotoxicity assay using WST-1. Values are means from two independent experiments performed with each condition in triplicate; bars indicate SD.

Article Snippet: Parental HEK293 cells (HEK; ATCC, Manassas, VA) and established human pancreatic carcinoma cell lines (Capan-1, Capan-2, PANC-1, AsPC-1, BxPC-3, MiaPaCa-2, and PaTu-8902 (PaTu)) [ 36 ] were cultured in Dulbecco modified Eagle medium/Ham F-12 medium with l -glutamine (PAA, Pasching, Austria) and 10% fetal bovine serum (Invitrogen, Carlsbad, CA), gentamicin (50 μg/ml), and amphothericin B (0.25 μg/ml; PAA); in addition, the culture medium of MRP5-transfected, MRP5-overexpressing HEK293 cells (HEK/MRP5 cells; Millennium Pharmaceuticals, Cambridge, MA) contained geneticin (250 μg/ml).

Techniques: Cytotoxicity Assay

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: ITGB6 shows specificity and upregulation in human cancers. (A) Human Protein Atlas analysis of RNA expression of ITGB6 across various cancer types. Normalized RNA expression levels are given in fragments per kilobase million (FPKM). Data are acquired from TCGA. Graphs show the mean and SD. (B) Differential gene expression analysis of ITGB6 across tumor and normal tissue. Data are obtained from a standardized concatenated data set from GEO, GTex, TCGA, and TARGET databases. RNA-seq tumor and normal tissue samples are obtained from the same patients and from adjacent sites. Graphs were generated using the TNMplot tool and were normalized using DESeq2. Fold-change (FC) of medians and Mann-Whitney test p -values are shown.

Article Snippet: ITGB6 knockout induces T-cell killing of HNSCC and PAAD cells. (A) ITGB6 RNA of selected human cancer cell lines from the Human Protein Atlas. (B) Flow cytometry analysis of the percentage of cancer cells that are ITGB6+ compared to unstained controls. (C) Western blot validation of the CRISPR knockout of ITGB6 in FaDu, CAL27, and Capan-2 cells. (D) Colony formation assay of CRISPR control (CTRL) cells and ITGB6 knockouts (KO-1) for FaDu and CAL27 cells (n = 3). (E) Quantification of colony formation assay.

Techniques: RNA Expression, Gene Expression, RNA Sequencing, Generated, MANN-WHITNEY

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: ITGB6 knockout induces T-cell killing of HNSCC and PAAD cells. (A) ITGB6 RNA of selected human cancer cell lines from the Human Protein Atlas. (B) Flow cytometry analysis of the percentage of cancer cells that are ITGB6+ compared to unstained controls. (C) Western blot validation of the CRISPR knockout of ITGB6 in FaDu, CAL27, and Capan-2 cells. (D) Colony formation assay of CRISPR control (CTRL) cells and ITGB6 knockouts (KO-1) for FaDu and CAL27 cells (n = 3). (E) Quantification of colony formation assay. Statistical analysis was performed using an unpaired t-test. Randomly chosen fields of view from the co-culture of FaDu (F), CAL-27 (G), or Capan-2 (H) cells (green) and TALL-104 T-cells (blue) (n = 10). Cells were pretreated for 8 hrs with latent TGFβ. Quantification of cancer cell counts for FaDu (I), CAL27 (M), and Capan-2 (Q). Quantification of the percentage of cancer cells that are dead for FaDu (J), CAL27 (N), and Capan-2 (R). Quantification of TALL-104 T-cell counts for the FaDu cohort (K), the CAL27 cohort (O), and the Capan-2 cohort (S). Quantification of the percentage of T cells that are dead for the FaDu cohort (L), the CAL27 cohort (P), and the Capan-2 cohort (T). One-way ANOVA: P < 0.0001 (****), P < 0.01 (**).

Article Snippet: ITGB6 knockout induces T-cell killing of HNSCC and PAAD cells. (A) ITGB6 RNA of selected human cancer cell lines from the Human Protein Atlas. (B) Flow cytometry analysis of the percentage of cancer cells that are ITGB6+ compared to unstained controls. (C) Western blot validation of the CRISPR knockout of ITGB6 in FaDu, CAL27, and Capan-2 cells. (D) Colony formation assay of CRISPR control (CTRL) cells and ITGB6 knockouts (KO-1) for FaDu and CAL27 cells (n = 3). (E) Quantification of colony formation assay.

Techniques: Knock-Out, Flow Cytometry, Western Blot, Biomarker Discovery, CRISPR, Colony Assay, Control, Co-Culture Assay

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: Suppressed growth of ITGB6 -knockout tumors in immunocompetent mice. (A) Flow cytometry analysis of the percentage of mouse cancer cells that are ITGB6+ compared to unstained controls. (B) Western blot validation of CRISPR knockdown of ITGB6 in MOC1 and KPCY cells. (C) Colony formation assay of CRISPR control (CTRL) cells and ITGB6-knockout (KO-1, KO-2) cells for MOC1 and KPCY cells. (D) Quantification of colony formation assay. Statistical analysis was performed using one-way ANOVA: P < 0.05 (*). (E) Experimental timeline for C57BL/6 mice injected with syngeneic tumor cells. Tumor volume of mice starting upon tumor formation at day 5 for MOC1 (F) and KPCY (H) cohorts (n = 10). Mass of MOC1 (G) and KPCY (I) tumors harvested at day 25. Statistical comparison between CTRL and KO-1 cohorts was performed using an unpaired t-test: P < 0.001 (***), P < 0.05 (*). (J) Heat map of Luminex panel showing fold change of cytokines of tumor interstitial fluid harvested from MOC1 and KPCY tumors (n = 5). Red indicates upregulation and green downregulation of cytokines in ITGB6 KO tumors compared to control tumors. (K) Concentrations of select tumor interstitial fluid cytokines from MOC1 and KPCY (L) as measured by the Luminex panel. Statistical comparison between CTRL and KO-1 cohorts was performed using an unpaired t-test. Representative 10× images of MOC1 (M) and KPCY (O) from scanned IHC slides showing CD8 T-cell infiltration across control and ITGB6-deficient cells. Quantification of CD8-stained IHC slides for MOC1 (N) (control: n = 5, KO: n = 10) and KPCY (P) (control: n = 7, KO: n = 8) cohorts, showing p -values calculated using an unpaired t-test.

Article Snippet: ITGB6 knockout induces T-cell killing of HNSCC and PAAD cells. (A) ITGB6 RNA of selected human cancer cell lines from the Human Protein Atlas. (B) Flow cytometry analysis of the percentage of cancer cells that are ITGB6+ compared to unstained controls. (C) Western blot validation of the CRISPR knockout of ITGB6 in FaDu, CAL27, and Capan-2 cells. (D) Colony formation assay of CRISPR control (CTRL) cells and ITGB6 knockouts (KO-1) for FaDu and CAL27 cells (n = 3). (E) Quantification of colony formation assay.

Techniques: Knock-Out, Flow Cytometry, Western Blot, Biomarker Discovery, CRISPR, Knockdown, Colony Assay, Control, Injection, Comparison, Luminex, Staining

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: ITGB6 decreases overall survival in patients. Curves show the overall survival of patients with various cancer types stratified based on ITGB6 expression. Patients with high ITGB6 have expression levels above +0.25 σ of the mean, and ITGB6 low patients have expression below -0.25 σ. The graphs were generated using TCGA data through the cBioPortal. Statistical analysis of Kaplan-Meier curves and corresponding hazard ratios (HR) was performed using the Log-rank test.

Article Snippet: ITGB6 knockout induces T-cell killing of HNSCC and PAAD cells. (A) ITGB6 RNA of selected human cancer cell lines from the Human Protein Atlas. (B) Flow cytometry analysis of the percentage of cancer cells that are ITGB6+ compared to unstained controls. (C) Western blot validation of the CRISPR knockout of ITGB6 in FaDu, CAL27, and Capan-2 cells. (D) Colony formation assay of CRISPR control (CTRL) cells and ITGB6 knockouts (KO-1) for FaDu and CAL27 cells (n = 3). (E) Quantification of colony formation assay.

Techniques: Expressing, Generated

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: Immune checkpoint blockade is modulated by ITGB6 expression. (A) Pan-cancer analysis of ITGB6 RNA expression level normalized using DESeq2 of patients who are either responsive or unresponsive to immune checkpoint blockade. RNA data are extracted from a TCGA data set of multiple cancer types of patients who underwent immune checkpoint blockade therapy. Graphs were generated using the ROCplot tool, and the results of an unpaired t-test are shown. (B) Kaplan-Meier curves of overall survival of patients undergoing immune checkpoint blockade therapy stratified into low and high ITGB6 expression about the mean. Statistical analysis was performed using the Log-rank test.

Article Snippet: ITGB6 knockout induces T-cell killing of HNSCC and PAAD cells. (A) ITGB6 RNA of selected human cancer cell lines from the Human Protein Atlas. (B) Flow cytometry analysis of the percentage of cancer cells that are ITGB6+ compared to unstained controls. (C) Western blot validation of the CRISPR knockout of ITGB6 in FaDu, CAL27, and Capan-2 cells. (D) Colony formation assay of CRISPR control (CTRL) cells and ITGB6 knockouts (KO-1) for FaDu and CAL27 cells (n = 3). (E) Quantification of colony formation assay.

Techniques: Expressing, RNA Expression, Generated