Journal: bioRxiv

Article Title: Metabolic and Anti-Proliferative Responses of Pancreatic Cancer Cells to Ultrasound and Nanobubble Treatment

doi: 10.64898/2026.04.24.720507

Figure Lengend Snippet: Passive cavitation detection spectrograms recorded during US exposure of pancreatic cancer cells treated with NBs. US was applied either immediately after NB addition (t = 0, extracellular NBs present) or after a 1-hour incubation followed by washing to remove extracellular NBs (1 h wash). Spectrograms show acoustic emissions for PANC-1 and BxPC-3 cells under both conditions. Immediate US exposure produced stronger broadband and harmonic emissions, whereas delayed US after washing resulted in reduced acoustic signal intensity, indicating reduced cavitation activity when extracellular NBs were removed and primarily cell-associated NBs remained (Supplemental Video1)

Article Snippet: BxPC-3 cells (ATCC CRL-1687) were maintained in RPMI 1640 (GibcoTM, Thermo Fisher Scientific, Milano, cat# 11875093) supplemented with 10% (v/v) FCS (GibcoTM, Thermo Fisher Scientific, Milano, cat# 10437-036) under identical incubation conditions.

Techniques: Incubation, Produced, Activity Assay

Journal: bioRxiv

Article Title: Metabolic and Anti-Proliferative Responses of Pancreatic Cancer Cells to Ultrasound and Nanobubble Treatment

doi: 10.64898/2026.04.24.720507

Figure Lengend Snippet: Cell viability measured using trypan blue exclusion in BxPC-3 and PANC-1 cells at 1 hour and 24 hours after treatment. Treatment groups included untreated control, US only, US applied immediately after NB addition (NB + US t = 0), US applied after 1-hour NB incubation followed by washing (NB 1 h + US), and NBs only (NB 1 h). Immediate US exposure in the presence of extracellular NBs resulted in a greater reduction in viability compared with delayed US exposure after washing. Viability differences were more pronounced at 24 hours than at 1 hour, indicating delayed treatment effects. Data are presented as percent viability relative to untreated control. Statistical significance is indicated, n=8, (*) P < 0.05, (**) P < 0.01.

Article Snippet: BxPC-3 cells (ATCC CRL-1687) were maintained in RPMI 1640 (GibcoTM, Thermo Fisher Scientific, Milano, cat# 11875093) supplemented with 10% (v/v) FCS (GibcoTM, Thermo Fisher Scientific, Milano, cat# 10437-036) under identical incubation conditions.

Techniques: Control, Incubation

Journal: bioRxiv

Article Title: Metabolic and Anti-Proliferative Responses of Pancreatic Cancer Cells to Ultrasound and Nanobubble Treatment

doi: 10.64898/2026.04.24.720507

Figure Lengend Snippet: (a) Representative immunofluorescence images of BxPC-3 cells stained for Ki-67 (proliferation marker) and nuclei (DAPI) under different treatment conditions: untreated, US only, US applied immediately after NB addition (NB + US 0T), US applied after 1-hour NB incubation and washing (NB + US 1T), and NBs only. (b) Quantification of the percentage of Ki-67 positive proliferating cells for each treatment group. US combined with NBs significantly reduced proliferative activity compared with controls, with the greatest reduction observed when US was applied immediately after NB addition. Data are presented as mean ± SD with statistical significance indicated. (*) P < 0.05, (**) P < 0.01, (***) P < 0.001.

Article Snippet: BxPC-3 cells (ATCC CRL-1687) were maintained in RPMI 1640 (GibcoTM, Thermo Fisher Scientific, Milano, cat# 11875093) supplemented with 10% (v/v) FCS (GibcoTM, Thermo Fisher Scientific, Milano, cat# 10437-036) under identical incubation conditions.

Techniques: Immunofluorescence, Staining, Marker, Incubation, Activity Assay

Journal: bioRxiv

Article Title: Metabolic and Anti-Proliferative Responses of Pancreatic Cancer Cells to Ultrasound and Nanobubble Treatment

doi: 10.64898/2026.04.24.720507

Figure Lengend Snippet: (a-b) Optical density measurements over 72 hours representing metabolic activity measured using the MTT assay in BxPC-3 (a) and PANC-1 (b) cells following treatment. (c-d) Extracellular acidification rate (ECAR) measurements over time for BxPC-3 (c) and PANC-1 (d) cells, indicating glycolytic activity. (e-f) Oxygen consumption rate (OCR) measurements over time for BxPC-3 (e) and PANC-1 (f) cells, indicating mitochondrial respiration. Treatment groups included untreated control, US only, US applied immediately after NB addition, US applied after 1-hour NB incubation and washing, and NBs only. The data demonstrate intrinsic metabolic differences between cell lines and timing-dependent metabolic responses to US-NB treatments. Data were analyzed using GraphPad Prism 8. Mean ± SD is reported with two-way ANOVA with Tukey’s (*) P < 0.05, (**) P < 0.01, (***) P < 0.001, (****) P < 0.0001.

Article Snippet: BxPC-3 cells (ATCC CRL-1687) were maintained in RPMI 1640 (GibcoTM, Thermo Fisher Scientific, Milano, cat# 11875093) supplemented with 10% (v/v) FCS (GibcoTM, Thermo Fisher Scientific, Milano, cat# 10437-036) under identical incubation conditions.

Techniques: Activity Assay, MTT Assay, Control, Incubation

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