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

Journal: Mediators of Inflammation

Article Title: The Vitamin D3 Analog Calcipotriol Attenuates Pancreatic Cancer Malignancy via Downregulating Thrombospondin 1 in Pancreatic Stellate Cells

doi: 10.1155/mi/2632235

Figure Lengend Snippet: Effects of Cal on aPSC activation. (A) (Left) VDR mRNA expression in PDAC cell lines (AsPC‐1, MIA PaCa‐2, and PANC‐1) and aPSCs was determined by qRT‐PCR ( n = 3). (Right) CYP24A1 mRNA expression in PDAC or aPSCs treated with DMSO or Cal (100 nM and 48 h) was examined by qRT‐PCR ( n = 3). (B) VDR protein expression in PDAC cell lines (AsPC‐1, MIA PaCa‐2, and PANC‐1) and aPSCs was determined by western blot ( n = 3). (C) Correlation analysis between α‐SMA and VDR mRNA expression in aPSCs, with GAPDH normalization ( n = 9). (D) VDR and α‐SMA gene expression in aPSCs treated with DMSO or Cal (100 nM and 48 h) was evaluated by qRT‐PCR ( n = 3). (E) VDR and α‐SMA protein expression in aPSCs treated with DMSO or Cal (100 nM and 48 h) ( n = 4). (F) Immunocytochemistry showing α‐SMA expression in aPSCs treated with DMSO or Cal (100 nM and 48 hr) ( n = 3). (G) EZ4U assay indicating the impacts of Cal on the proliferation of aPSCs ( n = 3). (H) Transwell migration assay and (I) wound healing showing the effects of Cal on aPSCs’ migration ability ( n = 3). caPSCs, PSCs derived from pancreatic cancer; cpPSCs, PSCs derived from chronic pancreatitis; cuPSCs, culture‐activated PSCs derived from normal tissue; aPSCs, activated PSCs; HPF, high‐power field; Ctr, control group treated with DMSO. All experiments were conducted in triplicate. ns, not significant. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Human PDAC cell lines PANC‐1 (male, American Type Culture Collection [ATCC] CRL‐1469, RRID: CVCL_0480), MIA PaCa‐2 (male, ATCC CRL‐1420, RRID: CVCL_0428), and AsPC‐1 (female, ATCC CRL‐1682, RRID: CVCL_0152) were purchased directly from the ATCC (Manassas, VA, USA) in 2015.

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Western Blot, Gene Expression, Immunocytochemistry, Transwell Migration Assay, Migration, Derivative Assay, Control

Journal: Mediators of Inflammation

Article Title: The Vitamin D3 Analog Calcipotriol Attenuates Pancreatic Cancer Malignancy via Downregulating Thrombospondin 1 in Pancreatic Stellate Cells

doi: 10.1155/mi/2632235

Figure Lengend Snippet: Dose‐dependent effect of rTHBS1 on PDAC malignancy. (A) Transwell migration assays showing the response of PDAC cell lines PANC‐1 and MIA PaCa‐2 to varying concentrations of rTHBS1 (0, 0.5, and 5 μg/mL). (B) Transwell invasion assays were used to quantify the invasive potential of the same PDAC cell lines under the same rTHBS1 treatments. (C) Proliferation of PDAC cells was assessed by EZ4U assay after treatment with rTHBS1 at 0, 0.5, 5, and 20 μg/mL. (D) Wound healing assays complement the migration analysis, with images and quantification of the migration area closure. (E) Representative micrographs depicting morphological alterations in PANC‐1 and MIA PaCa‐2 cells when cultured in standard medium, aPSCs‐CM, and standard medium supplemented with 5 μg/mL of rTHBS1. All experiments were conducted in triplicate. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Human PDAC cell lines PANC‐1 (male, American Type Culture Collection [ATCC] CRL‐1469, RRID: CVCL_0480), MIA PaCa‐2 (male, ATCC CRL‐1420, RRID: CVCL_0428), and AsPC‐1 (female, ATCC CRL‐1682, RRID: CVCL_0152) were purchased directly from the ATCC (Manassas, VA, USA) in 2015.

Techniques: Migration, Cell Culture

Journal: Mediators of Inflammation

Article Title: The Vitamin D3 Analog Calcipotriol Attenuates Pancreatic Cancer Malignancy via Downregulating Thrombospondin 1 in Pancreatic Stellate Cells

doi: 10.1155/mi/2632235

Figure Lengend Snippet: Inhibition of aPSCs‐CM–driven malignancy in PDAC by THBS1 neutralizing antibody. THBS1 neutralizing Ab diminished aPSCs‐CM–induced migration (A, C, D), invasion (B), proliferation (E–F), and EMT (G–H) of PDAC but had no effects on Cal‐aPSCs‐CM–induced malignancy of PDAC. aPSCs‐CM, CM from aPSCs pretreated with DMSO; Cal‐aPSCs‐CM, CM harvested from aPSCs pretreated with 100 nM Cal for 48 h. CM was then pretreated with 1 μg/mL of THBS1 Ab or control IgG and added to the PDAC. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Human PDAC cell lines PANC‐1 (male, American Type Culture Collection [ATCC] CRL‐1469, RRID: CVCL_0480), MIA PaCa‐2 (male, ATCC CRL‐1420, RRID: CVCL_0428), and AsPC‐1 (female, ATCC CRL‐1682, RRID: CVCL_0152) were purchased directly from the ATCC (Manassas, VA, USA) in 2015.

Techniques: Inhibition, Migration, Control

Journal: Mediators of Inflammation

Article Title: The Vitamin D3 Analog Calcipotriol Attenuates Pancreatic Cancer Malignancy via Downregulating Thrombospondin 1 in Pancreatic Stellate Cells

doi: 10.1155/mi/2632235

Figure Lengend Snippet: Attenuation of aPSCs‐CM–induced PDAC aggressiveness by CD47 blockade. CD47 blocking Ab diminished aPSCs‐CM–induced migration (A, C, D), invasion (B), proliferation (E–F), and EMT (G–H) of PDAC but had no effects on Cal‐aPSCs‐CM–induced aggressiveness of PDAC. PDAC were pretreated with 2 μg/mL CD47 blocking Ab or control IgG. All experiments were conducted in triplicate. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Human PDAC cell lines PANC‐1 (male, American Type Culture Collection [ATCC] CRL‐1469, RRID: CVCL_0480), MIA PaCa‐2 (male, ATCC CRL‐1420, RRID: CVCL_0428), and AsPC‐1 (female, ATCC CRL‐1682, RRID: CVCL_0152) were purchased directly from the ATCC (Manassas, VA, USA) in 2015.

Techniques: Blocking Assay, Migration, Control

Journal: Mediators of Inflammation

Article Title: The Vitamin D3 Analog Calcipotriol Attenuates Pancreatic Cancer Malignancy via Downregulating Thrombospondin 1 in Pancreatic Stellate Cells

doi: 10.1155/mi/2632235

Figure Lengend Snippet: Differential impact on PDAC organoid morphology and EMT marker expression by aPSCs‐CM and antibody interventions. (A) Representative bright‐field images displaying PDAC organoids over 5 days in control (aPSCs‐CM), treated with Cal‐aPSCs‐CM, with THBS1 antibody‐depleted aPSCs‐CM, and with organoids where CD47 has been blocked, followed by treatment with aPSCs‐CM. (B) Western blot analysis of E‐cadherin and vimentin in organoids subjected to these varied treatments. (C) Protein expression quantification normalized to GAPDH, demonstrating the effect of THBS1 depletion and CD47 inhibition on EMT markers in PDAC organoids. All experiments were conducted in triplicate. ∗ p < 0.05 and ∗∗ p < 0.01. Scale bar: 100 μm.

Article Snippet: Human PDAC cell lines PANC‐1 (male, American Type Culture Collection [ATCC] CRL‐1469, RRID: CVCL_0480), MIA PaCa‐2 (male, ATCC CRL‐1420, RRID: CVCL_0428), and AsPC‐1 (female, ATCC CRL‐1682, RRID: CVCL_0152) were purchased directly from the ATCC (Manassas, VA, USA) in 2015.

Techniques: Marker, Expressing, Control, Western Blot, Inhibition

Journal: Small Science

Article Title: Ultrastructural Study of Microphysiological Systems of the Tumor Microenvironment

doi: 10.1002/smsc.202500567

Figure Lengend Snippet: Schematic overview of the study workflow. The MPS consisted of one‐chamber microfluidic devices used to generate two cancer‐on‐a‐chip models: PDAC and LAC. Cells were embedded in natural hydrogels composed of egg white (EW)‐gelatin and collagen type I, respectively. At the end of the culture period, devices were fixed, stained, dehydrated, disassembled, critically point‐dried, mounted, and coated for SEM and FIB‐SEM imaging. For LAC models, the FIB was additionally used to prepare thin lamellae for TEM. This workflow enabled high‐resolution observation of both external and internal organization of 3D multicellular structures, including cell–cell and cell–matrix interactions, matrix deposition, and intercellular communication.

Article Snippet: Human PDAC cell line PANC‐1, and human LAC cell line A549 from the American Type Culture Collection (ATCC, USA) were used for this work.

Techniques: Staining, Imaging

Journal: Small Science

Article Title: Ultrastructural Study of Microphysiological Systems of the Tumor Microenvironment

doi: 10.1002/smsc.202500567

Figure Lengend Snippet: Schematic of a generic MPS used in this study. The inner geometry consists of a single microfluidic channel with ports for hydrogel loading and perfusion of culture medium. The central chamber contains the 3D multicellular tumor culture embedded in a hydrogel matrix. Two distinct cancer‐on‐a‐chip models were implemented, each in a separate device: pancreatic ductal adenocarcinoma (PDAC, PANC‐1 cells) and lung adenocarcinoma (LAC, A549 cells). This schematic highlights the overall architecture of the chip and the spatial arrangement of the tumor constructs, hydrogel, and medium channels.

Article Snippet: Human PDAC cell line PANC‐1, and human LAC cell line A549 from the American Type Culture Collection (ATCC, USA) were used for this work.

Techniques: Construct

Journal: Small Science

Article Title: Ultrastructural Study of Microphysiological Systems of the Tumor Microenvironment

doi: 10.1002/smsc.202500567

Figure Lengend Snippet: (A) Scheme of the MPS of the TME of PDAC, consisting of a hydrogel‐based cancer‐on‐a‐chip model where the PANC‐1 cell line was used. (B) Brightfield microscopy representative images of the PANC‐1 aggregates grown over 14 days of culture in EW/gelatin hydrogels. Zoomed area shows single aggregate. Dual beam FIB‐SEM representative image of EW/gelatin hydrogel after 14 days of culture, displaying the typical nanoglobular morphology of these hydrogels. (C) Dual beam FIB‐SEM representative images of the surface of PANC‐1 aggregates in EW/gelatin hydrogel. ECM: extracellular matrix produced by the cells. Orange asterisk: EW/gelatin hydrogel. Green arrows: unions between cells. Orange arrows: unions between the cells and the hydrogel. Yellow arrows: spherical particles. Blue arrows: large spherical particles.

Article Snippet: Human PDAC cell line PANC‐1, and human LAC cell line A549 from the American Type Culture Collection (ATCC, USA) were used for this work.

Techniques: Microscopy, Produced

Journal: Small Science

Article Title: Ultrastructural Study of Microphysiological Systems of the Tumor Microenvironment

doi: 10.1002/smsc.202500567

Figure Lengend Snippet: (A) Dual beam FIB‐SEM representative images of the ECM secreted by the PANC‐1 cells. Image iii shows some measurements of the diameter of the fibers that comprise the ECM. (B) Measurements of the diameter and length of the ECM fibers secreted by the PANC‐1 cells. Data shown in violin plot as its distribution with median and the interquartile range (IQR), each dot represent a measurement ( n = 98 for fiber diameter and n = 46 for fiber length).

Article Snippet: Human PDAC cell line PANC‐1, and human LAC cell line A549 from the American Type Culture Collection (ATCC, USA) were used for this work.

Techniques:

Journal: Small Science

Article Title: Ultrastructural Study of Microphysiological Systems of the Tumor Microenvironment

doi: 10.1002/smsc.202500567

Figure Lengend Snippet: (A) Dual beam FIB‐SEM representative images of the internal cell organization of the aggregates formed by PANC‐1 cells in EW/gelatin hydrogels. Image i shows the surface of the aggregate milled with the ion beam along the yellow line. Images ii–v show internal sections of the aggregate shown in image i. Image vi belongs to a different aggregate. Individual cells are indicated by red numbers. For additional guidance to identify cell boundaries, Figure S1 provides the same images with a colored overlay mask highlighting the cellular regions. Light blue arrows: darker and denser intracellular areas. Green arrows: membrane projections connecting adjacent cells. Yellow arrows: EVs secreted by the cells. (B) Measurements of the size (i.e., diameter) of the EVs secreted by the PANC‐1 cells. Data shown in violin plot as its distribution with median and the IQR, each dot represents a measurement ( n = 48).

Article Snippet: Human PDAC cell line PANC‐1, and human LAC cell line A549 from the American Type Culture Collection (ATCC, USA) were used for this work.

Techniques: Membrane