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miapaca 2 cells  (ATCC)


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    Structured Review

    ATCC miapaca 2 cells
    Cytotoxicity and Cell adsorption property of PVA-U varying G.D. (a) IC 50 values <t>of</t> <t>MIAPaCa-2</t> cells treated with UDCA, PVA-U3, PVA-U15, and PVA-U25 for 24 h incubation at pH 7.4 and pH 6.5 (n = 3). (b) Cell viability of MIAPaCa-2 cells treated with 10 μg mL −1 PVA-U15 at pH 7.4 and pH 6.5 for 0, 4, 12, and 24 h incubation (n = 3). (c) Confocal images of MIAPaCa-2 cells treated with Cell tracker deep red and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 7.4 and pH 6.5 for 12 h incubation. Cell tracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 100 μm and 50 μm, respectively. Quantification of intracellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 24 h at pH 7.4 (d) and pH 6.5 (e) (n = 11). Quantification of pericellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 4 h at pH 7.4 (f) and pH 6.5 (g) (n = 11). (h) Confocal images of MIAPaCa-2 cells treated with lysotracker and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 6.5 for 4 h incubation. Lysotracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 50 μm and 10 μm, respectively. Statistical analysis was performed using unpaired two-tailed Student's t -test. Data are presented as mean ± S.D. ( N.S. : no significant difference, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).
    Miapaca 2 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1604 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Escape from cell uptake: Drug-Free cancer therapeutics regulated by hydrophobicity and negative charge"

    Article Title: Escape from cell uptake: Drug-Free cancer therapeutics regulated by hydrophobicity and negative charge

    Journal: Materials Today Bio

    doi: 10.1016/j.mtbio.2025.102752

    Cytotoxicity and Cell adsorption property of PVA-U varying G.D. (a) IC 50 values of MIAPaCa-2 cells treated with UDCA, PVA-U3, PVA-U15, and PVA-U25 for 24 h incubation at pH 7.4 and pH 6.5 (n = 3). (b) Cell viability of MIAPaCa-2 cells treated with 10 μg mL −1 PVA-U15 at pH 7.4 and pH 6.5 for 0, 4, 12, and 24 h incubation (n = 3). (c) Confocal images of MIAPaCa-2 cells treated with Cell tracker deep red and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 7.4 and pH 6.5 for 12 h incubation. Cell tracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 100 μm and 50 μm, respectively. Quantification of intracellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 24 h at pH 7.4 (d) and pH 6.5 (e) (n = 11). Quantification of pericellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 4 h at pH 7.4 (f) and pH 6.5 (g) (n = 11). (h) Confocal images of MIAPaCa-2 cells treated with lysotracker and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 6.5 for 4 h incubation. Lysotracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 50 μm and 10 μm, respectively. Statistical analysis was performed using unpaired two-tailed Student's t -test. Data are presented as mean ± S.D. ( N.S. : no significant difference, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).
    Figure Legend Snippet: Cytotoxicity and Cell adsorption property of PVA-U varying G.D. (a) IC 50 values of MIAPaCa-2 cells treated with UDCA, PVA-U3, PVA-U15, and PVA-U25 for 24 h incubation at pH 7.4 and pH 6.5 (n = 3). (b) Cell viability of MIAPaCa-2 cells treated with 10 μg mL −1 PVA-U15 at pH 7.4 and pH 6.5 for 0, 4, 12, and 24 h incubation (n = 3). (c) Confocal images of MIAPaCa-2 cells treated with Cell tracker deep red and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 7.4 and pH 6.5 for 12 h incubation. Cell tracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 100 μm and 50 μm, respectively. Quantification of intracellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 24 h at pH 7.4 (d) and pH 6.5 (e) (n = 11). Quantification of pericellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 4 h at pH 7.4 (f) and pH 6.5 (g) (n = 11). (h) Confocal images of MIAPaCa-2 cells treated with lysotracker and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 6.5 for 4 h incubation. Lysotracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 50 μm and 10 μm, respectively. Statistical analysis was performed using unpaired two-tailed Student's t -test. Data are presented as mean ± S.D. ( N.S. : no significant difference, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

    Techniques Used: Adsorption, Incubation, Fluorescence, Two Tailed Test

    Antitumor efficacy of PVA-U15. (a) Treatment schedule for MIAPaCa-2 tumors. BALB/c nude mice were inoculated with MIAPaCa-2 cells on day 0. The tumor-bearing mice were randomized and treated with UDCA (144 μM), PVA (10 μg mL −1 ) or PVA-U15 (10 μg mL −1 ). 100 μL of each solution was intratumorally injected 5 days per week from day 9 to day 21. (b) Tumor growth in mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). (c) Body weight of mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). Statistical analysis was performed using Tukey test. Data are presented as mean ± S.D. (∗ p < 0.05).
    Figure Legend Snippet: Antitumor efficacy of PVA-U15. (a) Treatment schedule for MIAPaCa-2 tumors. BALB/c nude mice were inoculated with MIAPaCa-2 cells on day 0. The tumor-bearing mice were randomized and treated with UDCA (144 μM), PVA (10 μg mL −1 ) or PVA-U15 (10 μg mL −1 ). 100 μL of each solution was intratumorally injected 5 days per week from day 9 to day 21. (b) Tumor growth in mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). (c) Body weight of mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). Statistical analysis was performed using Tukey test. Data are presented as mean ± S.D. (∗ p < 0.05).

    Techniques Used: Injection



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    Cytotoxicity and Cell adsorption property of PVA-U varying G.D. (a) IC 50 values <t>of</t> <t>MIAPaCa-2</t> cells treated with UDCA, PVA-U3, PVA-U15, and PVA-U25 for 24 h incubation at pH 7.4 and pH 6.5 (n = 3). (b) Cell viability of MIAPaCa-2 cells treated with 10 μg mL −1 PVA-U15 at pH 7.4 and pH 6.5 for 0, 4, 12, and 24 h incubation (n = 3). (c) Confocal images of MIAPaCa-2 cells treated with Cell tracker deep red and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 7.4 and pH 6.5 for 12 h incubation. Cell tracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 100 μm and 50 μm, respectively. Quantification of intracellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 24 h at pH 7.4 (d) and pH 6.5 (e) (n = 11). Quantification of pericellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 4 h at pH 7.4 (f) and pH 6.5 (g) (n = 11). (h) Confocal images of MIAPaCa-2 cells treated with lysotracker and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 6.5 for 4 h incubation. Lysotracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 50 μm and 10 μm, respectively. Statistical analysis was performed using unpaired two-tailed Student's t -test. Data are presented as mean ± S.D. ( N.S. : no significant difference, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).
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    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, <t>MIA</t> <t>PaCa-2</t> 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.
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    Image Search Results


    Cytotoxicity and Cell adsorption property of PVA-U varying G.D. (a) IC 50 values of MIAPaCa-2 cells treated with UDCA, PVA-U3, PVA-U15, and PVA-U25 for 24 h incubation at pH 7.4 and pH 6.5 (n = 3). (b) Cell viability of MIAPaCa-2 cells treated with 10 μg mL −1 PVA-U15 at pH 7.4 and pH 6.5 for 0, 4, 12, and 24 h incubation (n = 3). (c) Confocal images of MIAPaCa-2 cells treated with Cell tracker deep red and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 7.4 and pH 6.5 for 12 h incubation. Cell tracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 100 μm and 50 μm, respectively. Quantification of intracellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 24 h at pH 7.4 (d) and pH 6.5 (e) (n = 11). Quantification of pericellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 4 h at pH 7.4 (f) and pH 6.5 (g) (n = 11). (h) Confocal images of MIAPaCa-2 cells treated with lysotracker and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 6.5 for 4 h incubation. Lysotracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 50 μm and 10 μm, respectively. Statistical analysis was performed using unpaired two-tailed Student's t -test. Data are presented as mean ± S.D. ( N.S. : no significant difference, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

    Journal: Materials Today Bio

    Article Title: Escape from cell uptake: Drug-Free cancer therapeutics regulated by hydrophobicity and negative charge

    doi: 10.1016/j.mtbio.2025.102752

    Figure Lengend Snippet: Cytotoxicity and Cell adsorption property of PVA-U varying G.D. (a) IC 50 values of MIAPaCa-2 cells treated with UDCA, PVA-U3, PVA-U15, and PVA-U25 for 24 h incubation at pH 7.4 and pH 6.5 (n = 3). (b) Cell viability of MIAPaCa-2 cells treated with 10 μg mL −1 PVA-U15 at pH 7.4 and pH 6.5 for 0, 4, 12, and 24 h incubation (n = 3). (c) Confocal images of MIAPaCa-2 cells treated with Cell tracker deep red and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 7.4 and pH 6.5 for 12 h incubation. Cell tracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 100 μm and 50 μm, respectively. Quantification of intracellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 24 h at pH 7.4 (d) and pH 6.5 (e) (n = 11). Quantification of pericellular fluorescence intensity of MIAPaCa-2 cells treated with PVA-U0 (black), PVA-U3 (light blue), PVA-U15 (blue), and PVA-U25 (dark blue) incubated for 4 h at pH 7.4 (f) and pH 6.5 (g) (n = 11). (h) Confocal images of MIAPaCa-2 cells treated with lysotracker and 10 μg mL −1 PVA-U0-R, PVA-U3-R, PVA-U15-R, and PVA-U25-R at pH 6.5 for 4 h incubation. Lysotracker deep red and PVA-Us were shown in green and red color, respectively. Scale bars of images and enlarge images are 50 μm and 10 μm, respectively. Statistical analysis was performed using unpaired two-tailed Student's t -test. Data are presented as mean ± S.D. ( N.S. : no significant difference, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

    Article Snippet: MIAPaCa-2 cells, HT-29 cells, and A549 cells were purchased from the American Type Culture Collection (Manassas, VA, USA).

    Techniques: Adsorption, Incubation, Fluorescence, Two Tailed Test

    Antitumor efficacy of PVA-U15. (a) Treatment schedule for MIAPaCa-2 tumors. BALB/c nude mice were inoculated with MIAPaCa-2 cells on day 0. The tumor-bearing mice were randomized and treated with UDCA (144 μM), PVA (10 μg mL −1 ) or PVA-U15 (10 μg mL −1 ). 100 μL of each solution was intratumorally injected 5 days per week from day 9 to day 21. (b) Tumor growth in mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). (c) Body weight of mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). Statistical analysis was performed using Tukey test. Data are presented as mean ± S.D. (∗ p < 0.05).

    Journal: Materials Today Bio

    Article Title: Escape from cell uptake: Drug-Free cancer therapeutics regulated by hydrophobicity and negative charge

    doi: 10.1016/j.mtbio.2025.102752

    Figure Lengend Snippet: Antitumor efficacy of PVA-U15. (a) Treatment schedule for MIAPaCa-2 tumors. BALB/c nude mice were inoculated with MIAPaCa-2 cells on day 0. The tumor-bearing mice were randomized and treated with UDCA (144 μM), PVA (10 μg mL −1 ) or PVA-U15 (10 μg mL −1 ). 100 μL of each solution was intratumorally injected 5 days per week from day 9 to day 21. (b) Tumor growth in mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). (c) Body weight of mice treated with PVA (black), UDCA (blue), and PVA-U15 (red). Statistical analysis was performed using Tukey test. Data are presented as mean ± S.D. (∗ p < 0.05).

    Article Snippet: MIAPaCa-2 cells, HT-29 cells, and A549 cells were purchased from the American Type Culture Collection (Manassas, VA, USA).

    Techniques: Injection

    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.

    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

    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.

    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

    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.

    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

    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.

    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