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: Cancer Research Communications

Article Title: Alprazolam Reduces Inflammatory Cytokine Production in Pancreatic Cancer–Associated Fibroblasts

doi: 10.1158/2767-9764.CRC-25-0472

Figure Lengend Snippet: ALP alters the inflammatory secretome of PDAC CAFs in vitro . A, Annotated images of cytokine array membranes following incubation with conditioned media from C7-TA-PSC cells treated with 20 μmol/L ALP or DMSO for 24 hours. B, Quantification of the total pixel intensity for cytokines visualized in A . Data points reflect values of each technical duplicate from a single biological replicate ( n = 1). AU, arbitrary units. C, RT-qPCR quantification of the fold change in cytokine RNA expression from C7-TA-PSC cells after treatment with 20 μmol/L of each BZD relative to the DMSO control. Data were collected in technical duplicate and biological triplicate ( n = 3). D, Quantification of cytokine protein levels detected by individual ELISAs using the conditioned media from cells treated as in C . Data were collected in technical duplicate for three biological replicates ( n = 3). Data in B–D are represented as mean ± SD. All statistical analyses in C and D were conducted using one-way ANOVA with Dunnett correction for multiple comparisons. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: KPC1245 murine PDAC cells were derived from a Kras LSL-G12D/+ ; Trp53 LSL-R172H/+ ; Pdx1-Cre tumor in a female C57BL/6 mouse in the lab of Dr. David Tuveson (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

Techniques: In Vitro, Incubation, Quantitative RT-PCR, RNA Expression, Control

Journal: Cancer Research Communications

Article Title: Alprazolam Reduces Inflammatory Cytokine Production in Pancreatic Cancer–Associated Fibroblasts

doi: 10.1158/2767-9764.CRC-25-0472

Figure Lengend Snippet: ALP suppresses TLR4 signaling in PDAC CAFs. A, Heatmap of relative RNA expression levels for TLRs in a scRNA-seq profile of human fibroblasts from normal pancreas and PDAC tumor tissues analyzed in ref. . B, RT-qPCR validation of TLR4 RNA expression in C7-TA-PSC cells at baseline relative to GAPDH . C, Fold change in cytokine RNA expression levels determined by RT-qPCR following 24 hours of treatment with DMSO, 20 μmol/L ALP, and/or 2.5 μg/mL LPS. Experiments in B and C were conducted in technical and biological triplicate ( n = 3). Data are represented as mean ± SD. Statistical analyses in C were conducted using one-way ANOVA with a Tukey correction for multiple comparisons. ns, not significant; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: KPC1245 murine PDAC cells were derived from a Kras LSL-G12D/+ ; Trp53 LSL-R172H/+ ; Pdx1-Cre tumor in a female C57BL/6 mouse in the lab of Dr. David Tuveson (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

Techniques: RNA Expression, Quantitative RT-PCR, Biomarker Discovery

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

Journal: iScience

Article Title: Lomitapide mesylate and lomitapide target ALDOA to inhibit growth and enhance gemcitabine efficacy in PDAC

doi: 10.1016/j.isci.2026.115316

Figure Lengend Snippet: Lomitapide mesylate and lomitapide inhibit pancreatic ductal adenocarcinoma cell viability and proliferation (A) Statistical plots of high-throughput drug screening results from the FDA Drug Library for PDAC cell lines. Scatterplots show relative viability of BxPC3 (left) and SW1990 (right) cells after 72 h treatment with 884 FDA-approved drugs (10 μM, n = 3). Red dot indicates lomitapide mesylate and green dot indicates lomitapide. (B) CCK-8 assay showing cell viability of PDAC cells after compound treatment. Bar graphs represent relative viability of cells treated with 10 μM lomitapide mesylate, 10 μM lomitapide, or DMSO (vehicle control) for 24 h, n = 3. (C) Chemical structure of lomitapide mesylate. (D) Chemical structure of lomitapide. (E) Concentration-dependent inhibition of cell viability by lomitapide mesylate or lomitapide following 24 h treatment. (F) Time-dependent inhibition of cell viability by lomitapide mesylate or lomitapide at a concentration of 8 μM. (G and H) Inhibitory effects of lomitapide mesylate or lomitapide on the colony-forming capacity of PDAC cells following 6 h treatment at 8 μM. (H) shows the quantification of colony numbers in (G). Data represent mean ± SD of three independent experiments. Statistical significance was determined using an unpaired two-tailed Student’s t test. ∗, p < 0.05, ∗∗, p < 0.01, ∗∗∗, p < 0.001, ∗∗∗∗, p < 0.0001.

Article Snippet: The human PDAC cell lines BxPC3 and SW1990, and the human hepatocellular carcinoma cell line HepG2, were obtained from ATCC (USA).

Techniques: High Throughput Screening Assay, Drug discovery, CCK-8 Assay, Control, Concentration Assay, Inhibition, Two Tailed Test

Journal: iScience

Article Title: Lomitapide mesylate and lomitapide target ALDOA to inhibit growth and enhance gemcitabine efficacy in PDAC

doi: 10.1016/j.isci.2026.115316

Figure Lengend Snippet: Lomitapide mesylate and lomitapide induce G1 phase cell-cycle arrest and apoptosis in PDAC cells (A) Optical microscopy images showing vacuole formation induced by 8 μM lomitapide mesylate, or 8 μM lomitapide, or an equivalent volume of DMSO (vehicle control) for 6 h in BxPC3 and SW1990 cells. Scale bars, 20 μm (applies to all images in this panel). (B) Transmission electron microscopy analysis of lomitapide mesylate- and lomitapide-induced changes in the internal morphology of BxPC3 and SW1990 cells. Cells were treated with the indicated treatments for 6 h prior to analysis. Scale bars, 2 μm (applies to all images in this panel). (C) Flow cytometric analysis of the cell cycle in PDAC cells treated with 8 μM lomitapide mesylate, 8 μM lomitapide or an equivalent volume of DMSO (vehicle control). BxPC3 cells (2 × 10 5 cells/well) were treated for 6 h, while SW1990 cells (4.5 × 10 5 cells/well) were treated for 12 h. Images on the left show representative flow cytometry plots, and the right panel presents the statistical results of the percentage of cells in each cell cycle phase across each cell line. (D and E) Flow cytometric analysis of apoptosis in PDAC cells treated with the indicated treatments for 24 h. (E) shows the quantitative statistical results of total apoptotic rates. (F and G) Apoptotic analysis of BxPC3 cells treated with 8 μM lomitapide mesylate or 8 μM lomitapide at extended time points. (F) shows the quantitative statistical results of total apoptotic rates, while (G) presents representative annexin V-PE/7-AAD flow cytometry plots. The 0 h group corresponds to the drug-free blank control that is common and identical for lomitapide mesylate and lomitapide. Only one 0 h plot is shown for clarity, as the baseline was the same for both treatments. Statistical analyses were performed using Student’s t tests for two group’s comparisons and one-way ANOVA for multiple comparisons. Data represent mean ± SD of three independent experiments. ∗, p < 0.05, ∗∗, p < 0.01, ∗∗∗, p < 0.001, n.s., not significant.

Article Snippet: The human PDAC cell lines BxPC3 and SW1990, and the human hepatocellular carcinoma cell line HepG2, were obtained from ATCC (USA).

Techniques: Microscopy, Control, Transmission Assay, Electron Microscopy, Flow Cytometry

Journal: iScience

Article Title: Lomitapide mesylate and lomitapide target ALDOA to inhibit growth and enhance gemcitabine efficacy in PDAC

doi: 10.1016/j.isci.2026.115316

Figure Lengend Snippet: Lomitapide mesylate and lomitapide inhibit PDAC independently of lipid metabolism, autophagy suppression, and P38 signaling (A) MTTP mRNA expression in human tissues, as retrieved from The Human Protein Atlas database. (B) MTTP mRNA expression in human cancer cell lines, as retrieved from The Human Protein Atlas database. (C) Basal MTTP expression in HepG2, BxPC3, and SW1990 cells. (D) Oil Red O staining of BxPC3 and SW1990 cells treated with 8 μM lomitapide mesylate, or 8 μM lomitapide, or an equivalent volume of DMSO (vehicle control) for 6 h. Scale bars, 200 μm (applies to all images in [D]). (E) LC3B-II and p62 protein expression in BxPC3 and SW1990 cells following the indicated treatments. (F) LC3 transformation assay in cells following treatment with 8 μM lomitapide mesylate, or 8 μM lomitapide, or an equivalent volume of DMSO (vehicle control), in combination with autophagy inhibitors. Cells were pre-treated with autophagy inhibitors (CQ, 20 μM; NH 4 Cl, 20 mM; or E64D [10 μg/mL] + pepstatin A [10 μg/mL]) for 1 h, followed by treatment with the aforementioned agents for 6 h. Protein extracts were then analyzed for LC3B expression. (G and H) Monitoring autophagic flux in PDAC cells using the mRFP-GFP-LC3 dual-labeling system. BxPC3 and SW1990 cell lines with lentivirus-mediated stable overexpression of stubRFP-sensGFP-LC3 were constructed to track autophagic flux. Following the indicated treatments, the distribution of LC3-positive puncta was visualized via laser confocal microscopy. Yellow fluorescent spots (merged mRFP and GFP signals) represent autophagosomes, while red fluorescent spots (mRFP-only signals, due to GFP quenching in the acidic environment of autolysosomes) indicate autolysosomes. Statistical analysis of the percentages of yellow and red puncta was performed to quantify changes in autophagic flux (H), n = 3. Scale bars, 20 μm (applies to all images in [G]). (I and J) Lomitapide mesylate and lomitapide were added 1 h after pretreatment with autophagy inhibitors or an activator, and cell viability was assessed 6 h thereafter. Autophagy inhibitors and activators used included WM, 5 μM; 3 MA, 5 mM; CQ, 20 μM; NH 4 Cl, 20 mM; E64D (10 μg/mL) + pepstatin A (10 μg/mL); or rapamycin, 10 μM ( n = 3). (K) BxPC3 and SW1990 cells were treated with the indicated treatments for 3 and 6 h, and the target proteins as well as their associated proteins were detected. (L) BxPC3 and SW1990 cells were pre-treated with SB202190 (10 μM) for 1 h, followed by the addition of the indicated treatments; cell viability was then assessed 6 h later ( n = 3). Data represent mean ± SD of three independent experiments. Statistical significance was determined using an unpaired two-tailed Student’s t test. ∗, p < 0.05, n.s., not significant.

Article Snippet: The human PDAC cell lines BxPC3 and SW1990, and the human hepatocellular carcinoma cell line HepG2, were obtained from ATCC (USA).

Techniques: Expressing, Staining, Control, Transformation Assay, Labeling, Over Expression, Construct, Confocal Microscopy, Two Tailed Test

Journal: iScience

Article Title: Lomitapide mesylate and lomitapide target ALDOA to inhibit growth and enhance gemcitabine efficacy in PDAC

doi: 10.1016/j.isci.2026.115316

Figure Lengend Snippet: ALDOA, A potential target molecule of lomitapide mesylate and lomitapide (A) DARTS samples from lomitapide-treated SW1990 cells were subjected to Coomassie Blue staining and silver staining. The red box marks the gel areas of control and experimental samples that were analyzed by mass spectrometry. (B) COG function classification of identified proteins. The vertical axis represents the number of marked proteins, and the horizontal axis shows different COG functional categories. (C) GO functional enrichment analysis results. (D and E) Molecular docking (MOE 2019) analyzed binding interactions and sites between lomitapide and ALDOA. (E) Binding mode of lomitapide (purple sticks) with ALDOA (ribbon model), with key interacting residues (Lys-229, Lys-107, Lys-146, Tyr-363 and Arg-148) labeled. The protein structure of ALDOA was retrieved from the RCSB website, with PDB ID: 2ALD. (F) DARTS validation of ALDOA as a target of lomitapide in SW1990 cells: SW1990 cell lysates were treated with 100 μM lomitapide, and the stability of the ALDOA protein was assessed. Pronase digestion was performed for 10 and 20 min, respectively. The increased stability of ALDOA in lomitapide-treated lysates indicates its interaction with lomitapide. (G) DARTS assay demonstrated dose-dependent lomitapide-ALDOA binding. SW1990 lysates were incubated with lomitapide (various concentrations, 1 h) and then digested with pronase (10 min). (H) ALDOA expression in BxPC3/SW1990 cells following treatment with the indicated treatments. (I) ALDOA enzymatic activity in BxPC3/SW1990 cells following the indicated treatments. (J) Boxplot showing ALDOA expression levels in PDAC (analyzed via GEPIA). The red asterisk indicates a statistically significant difference between groups. (K) Kaplan-Meier curve for overall survival of PDAC patients (from TCGA dataset) stratified by ALDOA .TPM expression levels (high vs. low). (L) Overall survival of pancreatic cancer patients (from the KM Plotter database) stratified by ALDOA .TPM expression levels. (M and N) Immunohistochemical (IHC) staining of ALDOA in 90 paired PDAC tumor tissues (left) and paratumor tissues (right) (M). ALDOA-positive signals (brownish-yellow staining) were markedly enriched in tumor tissues compared with paratumor tissues. (N) shows the quantitative statistical analysis of ALDOA IHC staining intensity. Scale bars, 200 μm (applies to all images in [M]). (O) ALDOA expression and survival in 90 paired PDAC patients. (P and Q) OCR in BxPC3/SW1990 cells following the indicated treatments for 6 h via Seahorse XF analyzer. (R and S) ECAR in BxPC3/SW1990 cells following the indicated treatments via Seahorse XF analyzer. (T) ATP levels in BxPC3/SW1990 cells following the indicated treatments for 3 h or 6 h. (U) Comparison of ALDOA protein levels between control and shRNA-mediated ALDOA -knockdown BxPC3/SW1990 cells. (V) Colony formation assay of BxPC3 and SW1990 cells with ALDOA knockdown. (W) Cell viability of BxPC3/SW1990 cells ( ALDOA -KD/Con) following the indicated treatments for 48 h. Data represent mean ± SD of three independent experiments. Statistical significance was determined using an unpaired two-tailed Student’s t test. ∗, p < 0.05, ∗∗, p < 0.01, ∗∗∗, p < 0.001, ∗∗∗∗, p < 0.0001, n.s., not significant.

Article Snippet: The human PDAC cell lines BxPC3 and SW1990, and the human hepatocellular carcinoma cell line HepG2, were obtained from ATCC (USA).

Techniques: Staining, Silver Staining, Control, Mass Spectrometry, Functional Assay, Binding Assay, Labeling, Biomarker Discovery, Incubation, Expressing, Activity Assay, Immunohistochemical staining, Immunohistochemistry, Comparison, shRNA, Knockdown, Colony Assay, Two Tailed Test

Journal: iScience

Article Title: Lomitapide mesylate and lomitapide target ALDOA to inhibit growth and enhance gemcitabine efficacy in PDAC

doi: 10.1016/j.isci.2026.115316

Figure Lengend Snippet: Lomitapide mesylate or lomitapide combined with gemcitabine yields superior outcomes (A) CCK-8 assay showing the viability of ALDOA -knockdown PDAC cells treated with gemcitabine for 48 h. (B–E) Dose-response matrices illustrating the combination effects of lomitapide mesylate/gemcitabine and lomitapide/gemcitabine in BxPC3 and SW1990 cells. (B) Dose-response matrix of lomitapide mesylate + gemcitabine in BxPC3 cells. (C) Dose-response matrix of lomitapide mesylate + gemcitabine in SW1990 cells. (D) Dose-response matrix of lomitapide + gemcitabine in BxPC3 cells. (E) Dose-response matrix of lomitapide + gemcitabine in SW1990 cells. For (B–E): BxPC3 (3 × 10 3 ) and SW1990 (5 × 10 3 ) cells were seeded in 96-well plates and incubated overnight. The following day, cells were treated with 6 × 6 matrix combinations of gemcitabine plus lomitapide mesylate or lomitapide at the indicated concentrations in a total volume of 100 μL for 48 h. Cell viability was then assessed using the CCK-8 assay. ZIP synergy scores were calculated via Synergy Finder, where a score > 10 indicates synergism, between −10 and 10 indicates additivity, and < −10 indicates antagonism.

Article Snippet: The human PDAC cell lines BxPC3 and SW1990, and the human hepatocellular carcinoma cell line HepG2, were obtained from ATCC (USA).

Techniques: CCK-8 Assay, Knockdown, Incubation