Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Low PAFAH1B1 level is closely associated with the radioresistance of 125 I particle radiation in PDAC cells. BxPC-3-RR, PANC-1-RR cells and their parental BxPC-3-P and PANC-1-P cells were exposed to 0, 2, 4 and 8 Gy radiation. ( A , C ) Colony formation in PDAC cells was monitored by colony formation assay. ( B , D ) Cell viability of PDAC cells was assessed by CCK-8 assay. BxPC-3-RR, PANC-1-RR cells and their parental BxPC-3-P and PANC-1-P cells were exposed to 4 Gy radiation. ( E ) Cell apoptosis was assessed by Annexin-V-APC/PI staining and flow cytometry analysis. ( F ) The mRNA levels of SMAD4 and PAFAH1B1 in PDAC cells was detected by qRT-PCR. ( G ) The protein levels of SMAD4 and PAFAH1B1 in PDAC cells were detected by western blot. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Colony Assay, CCK-8 Assay, Staining, Flow Cytometry, Analysis, Quantitative RT-PCR, Western Blot

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Reinforced PAFAH1B1 promotes the 125 I particle radiation sensitivity in irradiation-resistant PDAC cells. BxPC-3-RR and PANC-1-RR cells were transfected with vector alone or PAFAH1B1 overexpression construct. BxPC-3-P and PANC-1-P cells were stably transduced with AAV-shNC or AAV-shPAFAH1B1. ( A ) The mRNA level of PAFAH1B1 in PDAC cells was detected by qRT-PCR. ( B ) The protein level of PAFAH1B1 in PDAC cells was detected by western blot. ( C - D ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( E - G ) Colony formation in PDAC cells was monitored by colony formation assay. PDAC cells were subjected to overexpression or knockdown, followed by 4 Gy radiation. ( H ) Cell apoptosis was assessed by Annexin-V-APC/PI staining and flow cytometry analysis. ( I ) DNA damage was assessed by Comet assay. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Sensitivity, Irradiation, Transfection, Plasmid Preparation, Over Expression, Construct, Stable Transfection, Transduction, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Colony Assay, Knockdown, Staining, Flow Cytometry, Analysis, Single Cell Gel Electrophoresis

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Induction of ferroptosis contributes to the radiosensitivity of PDAC cells. BxPC-3-RR and PANC-1-RR cells were exposed to 4 Gy radiation. The intracellular ROS ( A ) and iron levels ( E ) were detected by DCFH-DA and FerroOrange staining, respectively. Green, DCFH-DA; Red, FerroOrange. Scale bar, 50 μm. ( B ) Lipid peroxidation was monitored by BODIPY 581/591 C11 staining and flow cytometry analysis. ( C - D ) The levels of MDA and GSH were detected using commercial kits. MDA or GSH level was normalized to total protein concentration determined by BCA protein assay. ( F ) Cell apoptosis was assessed by Annexin-V-APC/PI staining and flow cytometry analysis. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Staining, Flow Cytometry, Analysis, Protein Concentration, Bicinchoninic Acid Protein Assay

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: PAFAH1B1 weakens radioresistance by fostering radiation-induced ferroptosis. BxPC-3-P and PANC-1-P cells were stably transduced with AAV-shNC or AAV-shPAFAH1B1, and BxPC-3-RR and PANC-1-RR cells were stably transfected with vector alone and PAFAH1B1 overexpression plasmid. ( A ) Lipid peroxidation was monitored by BODIPY 581/591 C11 staining and flow cytometry analysis. ( B - C ) The levels of MDA and GSH were detected using commercial kits. MDA or GSH level was normalized to total protein concentration determined by BCA protein assay. ( D ) Intracellular iron was monitored by FerroOrange staining. Red, FerroOrange. Scale bar, 50 μm. ( E ) The protein levels of GPX4, SLC7A11 and ACSL4 in PDAC cells were detected by western blot. BxPC-3-RR and PANC-1-RR cells were transfected with vector alone or PAFAH1B1 overexpression construct in the absence or presence of Fer-1. ( F ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( G ) Colony formation in PDAC cells was monitored by colony formation assay. BxPC-3-P and PANC-1-P cells were stably transduced with AAV-shNC or AAV-shPAFAH1B1, followed by Erastin treatment. ( H ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( I ) Colony formation in PDAC cells was monitored by colony formation assay. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Stable Transfection, Transduction, Transfection, Plasmid Preparation, Over Expression, Staining, Flow Cytometry, Analysis, Protein Concentration, Bicinchoninic Acid Protein Assay, Western Blot, Construct, CCK-8 Assay, Colony Assay

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: PUM2 is identified as a key regulator for increasing PAFAH1B1 expression via directly binding to its 3′UTR. ( A ) The mRNA level of PUM2 in PDAC tumors (n = 74) and matched adjacent normal pancreatic tissues (n = 74) was detected by qRT-PCR. ( B ) The correlation between PUM2 and PAFAH1B1 was analyzed by Spearman correlation analysis. ( C ) The mRNA level of PUM2 in BxPC-3-RR, PANC-1-RR and their parental cells was detected by qRT-PCR. BxPC-3-RR and PANC-1-RR cells were transduced with Lenti-NC or Lenti-PUM2. The mRNA levels of PUM2 ( D ) and PAFAH1B1 ( E ) were detected by qRT-PCR. ( F ) The protein levels of PUM2 and PAFAH1B1 were detected by western blot. ( G ) The mRNA stability of PAFAH1B1 in BxPC-3-RR and PANC-1-RR cells was monitored by RNA stability assay. ( H ) The luciferase activities of PAFAH1B1 promoter was measured. ( I ) 5’ UTR, CDS and 3’ UTR in BxPC-3-RR and PANC-1-RR cells were detected by dual luciferase reporter assay. ( J ) The interaction between PUM2 and PAFAH1B1 3’ UTR was assessed by RIP assay. Normal IgG served as a negative control. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Analysis, Transduction, Western Blot, Stability, Luciferase, Negative Control

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Overexpression of PUM2 competitively binds to PAFAH1B1 3′ UTR with miR-421 and miR-301a-3p. ( A ) Bioinformatics analysis predicted the putative binding sites between miR-421/miR-301-3p and PAFAH1B1 3’UTR. HKE293T cells were co-transfected with Luc-S-3’ UTR and mimics control (NC)/miR-421 mimics/miR-301a-3p mimics. ( B ) The luciferase activity of Luc-S-3’ UTR was detected by dual luciferase reporter assay. ( C ) The enrichments of miR-421 and miR-301a-3p were detected by Ago2-RIP assay. Normal IgG served as a negative control. HKE293T cells were transfected with miRNA mimics or inhibitor. ( D ) The mRNA level of PAFAH1B1 in BxPC-3-RR cells was detected by qRT-PCR. ( E ) The protein level of PAFAH1B1 in BxPC-3-RR cells was detected by western blot. Lentivirus-transduced PANC-1-RR cells were transfected with mimics control (NC)/miR-421 mimics/miR-301a-3p mimics. ( F ) The mRNA level of PAFAH1B1 was detected by qRT-PCR. ( G ) The protein level of PAFAH1B1 was detected by western blot. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Over Expression, Analysis, Binding Assay, Transfection, Control, Luciferase, Activity Assay, Negative Control, Quantitative RT-PCR, Western Blot

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: PUM2 enhances radiation-induced ferroptosis in a PAFAH1B1-dependent manner. BxPC-3-RR and PANC-1-RR cells were transduced with lentivirus and AAV. ( A ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( B ) Colony formation in PDAC cells was monitored by colony formation assay. ( C - D ) The levels of MDA and GSH were detected using commercial kits. MDA or GSH level was normalized to total protein concentration determined by BCA protein assay. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Transduction, CCK-8 Assay, Colony Assay, Protein Concentration, Bicinchoninic Acid Protein Assay

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Glycolysis inhibitors diminish the virus sensitivity of glycolytic PDAC cells MIA PaCa-2 and PK-59 cells were treated with SCH772984 (SCH) (200 nM) or 2DG (2 mM), followed by infection with OBP-401 (100 MOI) or OBP-702 (10 MOI). (A) Lactate secretion by MIA PaCa-2 and PK-45H cells treated with SCH772984 or 2DG, presented as fold-increase compared with the control group, which was set as 1.0. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (B) Cell lysates of MIA PaCa-2 and PK-45H cells treated with SCH or 2DG for 48 h were subjected to western blot analysis for ERK1/2, GLUT1, and LDHA. (C) MIA PaCa-2 and PK-45H cells were treated with SCH or 2DG, followed by infection with OBP-401 (100 MOI) for 24 or 48 h. Upper panels show representative photographs of immunocytochemical staining for GFP in each group 48 h after infection. Scale bars, 500 μm. Lower graphs show the fluorescence intensity of GFP analyzed under fluorescence microscopy. Data are expressed as mean (SD) of independent experiment ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (D) MIA PaCa-2 and PK-45H cells were co-treated with OBP-702 and SCH772984 or 2DG at the indicated dose for 72 h. Cell viability was quantified using the XTT assay and calculated relative to the mock-infected group. Data are expressed as mean (SD) of independent experiment ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (E) Cell lysates of MIA PaCa-2 and PK-45H cells co-treated with SCH or 2DG and OBP-702 (10 MOI) for 48 h were subjected to western blot analysis for E1A, p53, PARP, and cleaved C-PARP. β-actin was assayed as a loading control. The expression level of each protein was calculated relative to that of mock-treated cells, which was set at 1.0. N.S., not significant; ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Virus, Infection, Control, Western Blot, Staining, Fluorescence, Microscopy, XTT Assay, Expressing

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: p53 activation modulates glutamine metabolism in PDAC cells (A) Glutamine consumption in PDAC cells, presented as fold-increase compared with PBS, which was set as 1.0. (B) Outline of glutamine metabolism, shown from glutamine uptake to α-KG production. (C) Lysates of PDAC cells were subjected to western blot analysis for GDH1/2, OGDH, and IDH1. (D) PDAC cells were infected with OBP-301 or OBP-702 at an MOI of 100 for 48 h. The amount of intracellular α-KG in PDAC cells is shown as fold-increase compared with the mock-infected group, which was set as 1.0. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (E) PDAC cells were infected with OBP-301 or OBP-702 at the indicated MOIs for 72 h. Cell lysates were subjected to western blot analysis for GDH1/2, OGDH, and IDH1. (F) MIA PaCa-2 and PK-59 cells were infected with DL312 or Adp53 at the indicated MOIs for 24 h. The amount of intracellular αKG in PDAC cells is presented as fold-increase compared with mock-infected control groups. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences among four groups was determined using one-way ANOVA followed by Turkey’s multiple comparison procedure. (G) MIA PaCa-2 and PK-59 cells were infected with DL312 or Adp53 at the indicated MOIs for 48 h. Cell lysates were subjected to western blot analysis for p53, GDH1/2, OGDH, and IDH1. β-Actin was assayed as a loading control. The expression level of each protein was calculated relative to that of MIAPaCa-2 cells or mock-treated cells, which was set at 1.0. ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activation Assay, Western Blot, Infection, Control, Comparison, Expressing

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Comparison of metabolic phenotypes and virus sensitivity in subcutaneous tumor models with glycolytic and non-glycolytic PDAC cells (A) Representative photographs of immunohistochemical staining for LDHA, GLUT1, and IDH1 in each group. Scale bars, 100 μm. (B) Expression levels of LDHA, GLUT1, and IDH1, calculated by dividing the DAB intensity by the number of cells in randomly selected fields. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (C) MIA PaCa-2 tumor-bearing mice received intratumoral injections of PBS (black arrows) or OBP-702 (green arrows) every other day for 3 cycles. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (D) PK-59 tumor-bearing mice received intratumoral injections of PBS (black arrows) or OBP-702 (orange arrows). The upper right photographs show tumor-bearing mice in the control and OBP-702-treated groups. The lower right photographs show tumors in the mock and OBP-702 groups. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Comparison, Virus, Immunohistochemical staining, Staining, Expressing, Control

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Investigation of the relationship between PET/CT metabolic parameters and glycolytic activity of PDAC tumors (A and B) PET/CT images of MIA PaCa-2 tumor (A) and PK-59 tumor (B). The upper left (a) shows the horizontal section, whereas the lower left (b) shows the sagittal section, and the right (c) shows the coronal section. Dotted circles indicate the tumor area. (C) Comparison of SUVmax values for MIA PaCa-2 and PK-59 tumors. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (D and E) Comparison of MTV (D) and TLG (E) values for MIA PaCa-2 and PK-59 tumors at the indicated thresholds. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (F and G) Scatter diagrams demonstrating correlations between expression of LDHA (F) or GLUT1 (G) and preoperative SUVmax (left), MTV (40%) (center), and TLG (40%) (right) values in patients with PDAC ( n = 30). The statistical significance of the correlations in the scatterplots was determined using Pearson’s correlation analysis. N.S., not significant; ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Positron Emission Tomography-Computed Tomography, Activity Assay, Comparison, Expressing