Journal: Oncology Reports

Article Title: Role of iron and TfR1 in the application of high-dose ascorbate against pancreatic cancer

doi: 10.3892/or.2026.9083

Figure Lengend Snippet: Influence of high-dose ascorbate alone or in combination with ferric iron on the viability of human pancreatic cancer cell lines. The cell lines BxPC-3, MIA PaCa-2 and PANC-1 were either treated for 24 h with the indicated ascorbate concentrations alone (A), in the form of coincubation simultaneously with ascorbate and 100 µM FC (B), or incubated with 100 µM FC for 24 h as a preincubation before ascorbate treatment (C). Triton™ X-100 at 0.1% (v/v) served as positive control. Cell viability was measured after treatment by MUH assay. The results are presented as a percentage of fluorescence intensity relative to the untreated control. Three independent experiments were performed in duplicates. Error bars represent the mean ± SD, statistical analysis with one-way ANOVA and subsequent Dunnett's multiple comparisons test, confidence interval 95%. *P≤0.05, **P≤0.01, and ***P≤0.001. Asc, ascorbate; FC, ferric chloride; MUH, 4-methylumbelliferyl heptanoate.

Article Snippet: The human pancreatic carcinoma cell lines BxPC-3, MIA PaCa-2, and PANC-1 were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ).

Techniques: Incubation, Positive Control, Fluorescence, Control

Journal: Oncology Reports

Article Title: Role of iron and TfR1 in the application of high-dose ascorbate against pancreatic cancer

doi: 10.3892/or.2026.9083

Figure Lengend Snippet: Investigation of the effect of high-dose ascorbate treatment on different apoptosis markers in human pancreatic cancer cell lines. A possible induction of apoptosis in the human pancreatic cancer cell lines BxPC-3, MIA PaCa-2, and PANC-1 was investigated by testing caspase-3 cleavage by flow cytometry (A) and western blotting (B). Cells were treated for 6 h with the indicated ascorbate concentrations. 20 µM STS served as positive control. Three independent experiments were performed. Morphological nuclear changes were detected by fluorescence microscopy. The white scale bars represent 25 µm. (C). Cells were treated with the indicated ascorbate concentrations for 6 h and then fixed. 10 µM STS served as positive control. The nuclei were stained with DAPI (blue), the cytoskeleton with phalloidin (red). A representative experiment is shown. Cell cycle analysis was performed by flow cytometry (D). Cells were treated with the indicated ascorbate concentrations for 24 h, fixed, stained with PI, and subsequently detected. Treatment with 1 µM STS for 20 h served as positive control. Three independent experiments were performed. Error bars represent the mean ± SD, statistical analysis with one-way ANOVA and subsequent Dunnett's multiple comparisons test, confidence interval 95%. *P≤0.05 and ***P≤0.001. Asc, ascorbate; DAPI, diamidino-2-phenylindole; PI, propidium iodide; STS, staurosporine.

Article Snippet: The human pancreatic carcinoma cell lines BxPC-3, MIA PaCa-2, and PANC-1 were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ).

Techniques: Flow Cytometry, Western Blot, Positive Control, Fluorescence, Microscopy, Staining, Cell Cycle Assay

Journal: Oncology Reports

Article Title: Role of iron and TfR1 in the application of high-dose ascorbate against pancreatic cancer

doi: 10.3892/or.2026.9083

Figure Lengend Snippet: Investigation of the effect of high-dose ascorbate treatment on different ferroptosis markers in human pancreatic cancer cell lines. The human pancreatic cancer cell lines BxPC-3, MIA PaCa-2, and PANC-1 were treated with or without the ferroptosis inhibitor DFO for 24 h, after which cell viability was measured using the MUH assay (A). Triton™ X-100 at 0.1% (v/v) served as positive control. The results are presented as a percentage of the fluorescence intensity of the untreated control. Three independent experiments were performed in duplicates. As further signs of ferroptosis, protein expression of TfR1, GPX4, and LC3B-II was detected by western blotting (B) and quantified densitometrically (C). Cells were treated with the indicated ascorbate concentrations for 6 h, after which a western blot was performed. 5 µM RSL3 or 60 µM CQ together with 500 nM RAPA were used as positive controls. Signal intensity was analyzed densitometrically and normalized to GAPDH. One representative experiment out of two is shown. Error bars represent the mean ± SD, statistical analysis with one-way ANOVA and subsequent Dunnett's multiple comparisons test, confidence interval 95%. **P≤0.01 and ***P≤0.001. Asc, ascorbate; CQ, chloroquine; DFO, deferoxamine mesylate; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GPX4, glutathione peroxidase 4; MUH, 4-methylumbelliferyl heptanoate; RAPA, rapamycin; RSL3, RAS-selective lethal 3; TfR1, transferrin receptor 1.

Article Snippet: The human pancreatic carcinoma cell lines BxPC-3, MIA PaCa-2, and PANC-1 were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ).

Techniques: Positive Control, Fluorescence, Control, Expressing, Western Blot

Journal: Oncology Reports

Article Title: Role of iron and TfR1 in the application of high-dose ascorbate against pancreatic cancer

doi: 10.3892/or.2026.9083

Figure Lengend Snippet: The influence of iron and TfR1 on the ascorbate effect in human pancreatic cancer cell lines. To investigate the effect of iron on ascorbate action, the three human pancreatic cancer cell lines BxPC-3, MIA PaCa-2, and PANC-1 were either incubated simultaneously for 6 h with ascorbate at different concentrations and 100 µM ferric chloride (FC) (A) or treated with ascorbate after 24 h preincubation with ferric chloride (B). The intracellular ROS levels were determined after treatment by flow cytometry using the DCFH-DA assay. The percentage of DCF-positive cells is shown as a measure of intracellular ROS accumulation. Statistically significant differences between the combination treatment with iron and ascorbate treatment alone are marked by asterisks. Three independent experiments were performed. The intracellular LIP after 24-h incubation with ferric chloride was determined by flow cytometry using the calcein AM assay (C). The relative labile iron pool is shown in relation to the untreated control. Three independent experiments were performed. The basal protein expression of TfR1 in the pancreatic cancer cell lines and the non-malignant pancreatic ductal epithelial cell line HPDE6c7 was determined by western blotting, as well as the influence of ferric chloride on TfR1 expression in the three pancreatic cancer cell lines (D). The western blot results were also analyzed densitometrically. A representative experiment is shown for basal expression. For TfR1 expression after iron treatment, two independent experiments were performed, a representative western blot is shown. The influence of the 24-h ferric chloride treatment as well as ascorbate treatment was additionally determined at the mRNA level by qPCR (E). Error bars represent the mean ± SD, statistical analysis with one-way ANOVA and subsequent Dunnett's multiple comparisons test, confidence interval 95%. *P≤0.05, **P≤0.01, and ***P≤0.001. Asc, ascorbate; DCFH-DA, dichlorodihydrofluorescein diacetate; DCF, dichlorofluorescein; FC, ferric chloride; LIP, labile iron pool; ROS, reactive oxygen species; TfR1, transferrin receptor 1.

Article Snippet: The human pancreatic carcinoma cell lines BxPC-3, MIA PaCa-2, and PANC-1 were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ).

Techniques: Incubation, Flow Cytometry, DCFH-DA Assay, Calcein AM Assay, Control, Expressing, Western Blot

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: In vitro efficacy of paclitaxel and SB-Ts (A) Graph showing the cell viability (% of control cells) of Paca-44 and BxPC-3 cell line treated with paclitaxel (PTX), second- and third-generation of SB-Ts for 72 h. Table with IC 50 values as mean ± S.D. of three independent experiments presented. (B) Representative graphs depicting the effect of concentrations corresponding to IC 50 (5–30 nM) 100 nM and 300 nM of PTX, SB-T-121605, and SB-T-12606 on cellular proliferation in Paca-44 and BxPC-3 cells. The data are presented as mean ± standard deviation of two independent experiments.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: In Vitro, Control, Standard Deviation

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: Cell cycle changes after 24 h of incubation of cells with paclitaxel and experimental SB-Ts in vitro (A and B) Taxane concentrations corresponding to IC 50 values (5–30 nM) and 100 nM concentration were used in (A) Paca-44, (B) BxPC-3 cell line. Histograms display three independent experiments as mean ± SD.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Incubation, In Vitro, Concentration Assay

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: Western blot analysis of phospho-Bcl-2 (P-Bcl-2), PARP and cleaved caspase-3 expression in Paca-44 and BxPC-3 cells after paclitaxel and SB-Ts treatment (A) Cells treated with 25 nM (BxPC-3) or 30 nM (Paca-44) PTX, 5 nM SB-T-1216 (1216), 15 nM SB-T-121605 (121605), 5 nM SB-T-121606 (121606) or fresh medium without taxanes (CTRL) for 24 and 72 h. Expression of P-Bcl-2 (Ser70), PARP, and cleaved caspase-3 was analyzed employing western blot and relevant antibodies. Actin was used as a loading control. Data of one representative experiment out of three independent experiments shown. (B) Densitometric analysis of P-Bcl-2 (Ser70), cleaved form of PARP, and cleaved caspase-3. Data expressed as fold change of the respective control cells after 24 h and 72 h of treatment. The mean ± SD of three independent experiments shown. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Western Blot, Expressing, Control

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet: The analysis of cell death (A) Real-time monitoring by SYTOX Green with concentrations corresponding to IC 50 values (5–30 nM differing for each taxane-cell line combination), 100 and 300 nM PTX and SB-Ts. The data represent mean ± SD of three independent measurements. (B) The analysis of cell death by Annexin V and propidium iodide after 24, 48, and 72 h of incubation of cells with IC 50 and 100 nM concentrations of each taxane. All data presented as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (C) Caspase 3 and 7 activity after 24 and 72 h of Paca-44 and BxPC-3 cells incubation with PXT and SB-Ts. Data are displayed as % of caspase activity in control cells incubated with cultivation medium only. All data presented as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Incubation, Standard Deviation, Activity Assay, Control

Journal: iScience

Article Title: Third-generation taxanes SB-T-121605 and SB-T-121606 are effective in pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2024.109044

Figure Lengend Snippet:

Article Snippet: BxPC3 , Cell Lines Service GmbH Company, Eppelheim, Baden- Dr.-Eckener-Str. 8, Eppelheim, Baden-Württemberg, 69214, Germany , Product number: 305031.

Techniques: Recombinant

Journal: Cancer Science

Article Title: Tumor microenvironmental 15‐PGDH depletion promotes fibrotic tumor formation and angiogenesis in pancreatic cancer

doi: 10.1111/cas.15495

Figure Lengend Snippet: 15‐PGDH is expressed in the tumor microenvironment and promotes tumorigenesis of PDAC. (A) Representative immunofluorescence staining for 15‐PGDH, AE1/AE3, αSMA and CD45. The arrowheads show strongly 15‐PGDH‐positive cells. Scale bars, 100 μm. (B) Quantification of AE1/AE3 + , αSMA + and CD45 + cells. (C, D) 15‐pgdh expression in 15‐pgdh +/− mice and wild‐type mice was evaluated by qRT‐PCR (C) and western blotting (D). (E) Expression of 15‐pgdh in Panc02 cell lines and normal spleen tissue was evaluated by western blotting. (F) Strategy used to establish the orthotopic transplantation model. Panc02 cells deficient in 15‐pgdh were injected into the pancreas of wild‐type and 15‐pgdh +/− mice to generate orthotopic xenografts. After 21 days, the mice were euthanized, and the tumors were harvested and weighed. (G) Images showing pancreatic tumors and spleens. Scale bars, 1 cm. (H) Main tumor area of wild‐type and 15‐pgdh +/− mice measured by ImageJ software. (I) Volumes of tumors from wild‐type and 15‐pgdh +/− mice. (J) PGE2 expression was measured by LC/MS in wild‐type mice and 15‐pgdh +/− mice. NS, not significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The human PDAC cell lines PANC1, PK59, PK‐8, AsPC1, and MIAPaCa2 were obtained from the Japanese Collection of Research Bioresources Cell Bank and RIKEN Bioresource Center Cell Bank.

Techniques: Immunofluorescence, Staining, Expressing, Quantitative RT-PCR, Western Blot, Transplantation Assay, Injection, Software, Liquid Chromatography with Mass Spectroscopy

Journal: Cancer Science

Article Title: Tumor microenvironmental 15‐PGDH depletion promotes fibrotic tumor formation and angiogenesis in pancreatic cancer

doi: 10.1111/cas.15495

Figure Lengend Snippet: 15‐pgdh depletion in a highly fibrotic tumor microenvironment maintains angiogenesis levels. (A) Nano‐CT scanning of transplanted tumors from wild‐type and 15‐pgdh +/− mice and quantification of vessel branching. (B) Representative immunohistochemical staining and quantification of CD31 in wild‐type and 15‐pgdh +/− mouse tumors. Scale bars, 500 μm. (C) Expression of VEGFA in wild‐type and 15‐pgdh +/− mouse tumors was determined by qRT‐PCR. (D) Expression of VEGFA in five human PDAC cell lines and three CAF lines was determined by qRT‐PCR. (E) Morphology and quantification of branch points in HUVECs after culture in RPMI 1640 medium and CAF CM. Scale bars, 200 μm. (F) Morphology and quantification of branch points in HUVECs after culture in CAF CM and CAF CM + bevacizumab. Scale bars, 200 μm. (G) PK8 and PK8 cells + CAFs on CAM samples were evaluated for angiogenesis. Scale bars, 1 mm. (H) Immunofluorescence staining for VE‐cadherin and quantification of VE‐cadherin + cells. Scale bars, 200 μm. NS, not significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The human PDAC cell lines PANC1, PK59, PK‐8, AsPC1, and MIAPaCa2 were obtained from the Japanese Collection of Research Bioresources Cell Bank and RIKEN Bioresource Center Cell Bank.

Techniques: Immunohistochemical staining, Staining, Expressing, Quantitative RT-PCR, Immunofluorescence

Journal: Cancer Science

Article Title: Tumor microenvironmental 15‐PGDH depletion promotes fibrotic tumor formation and angiogenesis in pancreatic cancer

doi: 10.1111/cas.15495

Figure Lengend Snippet: Genetic deletion of 15‐pgdh favors fibrosis and angiogenesis in a PDAC mouse model. (A) Images showing pancreatic tumors and spleens of KC and PgKC mice. Main pancreas area of KC and PgKC mice measured by ImageJ. Scale bars, 1 cm. (B) H&E staining of KC and PgKC mice, Scale bars, 200 μm. (C) Representative Masson's trichrome staining of KC and PgKC mouse tumors. The graph on the right shows the area of collagen expression. KC: Kras LSL‐G12D/+ ; PgKC, Ptf1aCre /+ mice: 15pgdh −/− ; Kras LSL‐G12D/+ ; Ptf1aCre /+ mice. Scale bars, 500 μm. (D) Representative immunofluorescence staining for αSMA in KC and PgKC mice. The graph on the right shows the quantification of αSMA + cells. Scale bars, 50 μm. (E) Representative immunohistochemical staining for CD31 and quantification of CD31 + cells in KC and PgKC mice, Scale bars, 50 μm. NS, not significant; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The human PDAC cell lines PANC1, PK59, PK‐8, AsPC1, and MIAPaCa2 were obtained from the Japanese Collection of Research Bioresources Cell Bank and RIKEN Bioresource Center Cell Bank.

Techniques: Staining, Expressing, Immunofluorescence, Immunohistochemical staining