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Impact of Procyanidins from Different Berries on Caspase 8 Activation in Colon Cancer.

Minker C, Duban L, Karas D, Järvinen P, Lobstein A, Muller CD - Oxid Med Cell Longev (2015)

Bottom Line: Apoptosis induction is monitored by cell surface phosphatidylserine (PS) detection.Lowbush blueberry extract triggers the strongest activity.We highlight here that berries Pcys, especially lowbush blueberry Pcys, are of putative interest for nutritional chemoprevention of colorectal cancer in view of their apoptosis induction in a human colorectal cancer cell lines.

View Article: PubMed Central - PubMed

Affiliation: Laboratoire d'Innovation Thérapeutique, UMR 7200, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, BP 24, 67401 Illkirch Cedex, France.

ABSTRACT

Scope: The aim of this work is to identify which proapoptotic pathway is induced in human colon cancer cell lines, in contact with proanthocyanidins extracted from various berries.

Methods and results: Proanthocyanidins (Pcys) extracted from 11 berry species are monitored for proapoptotic activities on two related human colon cancer cell lines: SW480-TRAIL-sensitive and SW620-TRAIL-resistant. Apoptosis induction is monitored by cell surface phosphatidylserine (PS) detection. Lowbush blueberry extract triggers the strongest activity. When tested on the human monocytic cell line THP-1, blueberry Pcys are less effective for PS externalisation and DNA fragmentation is absent, highlighting a specificity of apoptosis induction in gut cells. In Pcys-treated gut cell lines, caspase 8 (apoptosis extrinsic pathway) but not caspase 9 (apoptosis intrinsic pathway) is activated after 3 hours through P38 phosphorylation (90 min), emphasizing the potency of lowbush blueberry Pcys to eradicate gut TRAIL-resistant cancer cells.

Conclusion: We highlight here that berries Pcys, especially lowbush blueberry Pcys, are of putative interest for nutritional chemoprevention of colorectal cancer in view of their apoptosis induction in a human colorectal cancer cell lines.

No MeSH data available.


Related in: MedlinePlus

Caspase 8 and P38 MAK pathways activation by lowbush blueberry proanthocyanidins in SW620 cells. (a) Caspase 8 activation curve responses were obtained with two fluorochrome-conjugated inhibitors of caspases consisting of a fluorophore (sulforhodamine for caspase 8 and carboxyfluorescein for caspase 9), a peptide specific for the active site of a particular caspase or many caspases, and a reactive functional group (fluoromethylketone or FMK). These inhibitors are cell permeable and noncytotoxic. Once inside the cell, the caspase inhibitors bind specifically through the peptide moiety to caspases that have been activated in apoptosis, and the FMK moiety covalently links the inhibitor to the caspase. The resulting signal is proportional to the number of active caspase enzymes that are present in the cell. (b) Directly conjugated phosphospecific antibodies were used to monitor the activation of P38 MAK and ATF2 pathways. All signals were monitored by capillary flow cytometry as described under M&M.
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fig6: Caspase 8 and P38 MAK pathways activation by lowbush blueberry proanthocyanidins in SW620 cells. (a) Caspase 8 activation curve responses were obtained with two fluorochrome-conjugated inhibitors of caspases consisting of a fluorophore (sulforhodamine for caspase 8 and carboxyfluorescein for caspase 9), a peptide specific for the active site of a particular caspase or many caspases, and a reactive functional group (fluoromethylketone or FMK). These inhibitors are cell permeable and noncytotoxic. Once inside the cell, the caspase inhibitors bind specifically through the peptide moiety to caspases that have been activated in apoptosis, and the FMK moiety covalently links the inhibitor to the caspase. The resulting signal is proportional to the number of active caspase enzymes that are present in the cell. (b) Directly conjugated phosphospecific antibodies were used to monitor the activation of P38 MAK and ATF2 pathways. All signals were monitored by capillary flow cytometry as described under M&M.

Mentions: After 24 hours, apple Pcys were shown to trigger activation of caspase 8 in both cell lines, but only caspase 9 in SW620 cells [54]. After 48 hours, the two caspases were fully activated. Our results here emphasize the fact that lowbush blueberry Pcys are more potent as they significantly activate both caspases 8 and 9 in SW620 (a) and SW480 (b) cell lines (Figure 6) after 24 and 48 hours. As for apple Pcys, shorter incubation times (i.e., 3 and 6 hours) resulted in caspase 8 activation only, highlighting the importance of the extrinsic apoptosis pathway.


Impact of Procyanidins from Different Berries on Caspase 8 Activation in Colon Cancer.

Minker C, Duban L, Karas D, Järvinen P, Lobstein A, Muller CD - Oxid Med Cell Longev (2015)

Caspase 8 and P38 MAK pathways activation by lowbush blueberry proanthocyanidins in SW620 cells. (a) Caspase 8 activation curve responses were obtained with two fluorochrome-conjugated inhibitors of caspases consisting of a fluorophore (sulforhodamine for caspase 8 and carboxyfluorescein for caspase 9), a peptide specific for the active site of a particular caspase or many caspases, and a reactive functional group (fluoromethylketone or FMK). These inhibitors are cell permeable and noncytotoxic. Once inside the cell, the caspase inhibitors bind specifically through the peptide moiety to caspases that have been activated in apoptosis, and the FMK moiety covalently links the inhibitor to the caspase. The resulting signal is proportional to the number of active caspase enzymes that are present in the cell. (b) Directly conjugated phosphospecific antibodies were used to monitor the activation of P38 MAK and ATF2 pathways. All signals were monitored by capillary flow cytometry as described under M&M.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4477188&req=5

fig6: Caspase 8 and P38 MAK pathways activation by lowbush blueberry proanthocyanidins in SW620 cells. (a) Caspase 8 activation curve responses were obtained with two fluorochrome-conjugated inhibitors of caspases consisting of a fluorophore (sulforhodamine for caspase 8 and carboxyfluorescein for caspase 9), a peptide specific for the active site of a particular caspase or many caspases, and a reactive functional group (fluoromethylketone or FMK). These inhibitors are cell permeable and noncytotoxic. Once inside the cell, the caspase inhibitors bind specifically through the peptide moiety to caspases that have been activated in apoptosis, and the FMK moiety covalently links the inhibitor to the caspase. The resulting signal is proportional to the number of active caspase enzymes that are present in the cell. (b) Directly conjugated phosphospecific antibodies were used to monitor the activation of P38 MAK and ATF2 pathways. All signals were monitored by capillary flow cytometry as described under M&M.
Mentions: After 24 hours, apple Pcys were shown to trigger activation of caspase 8 in both cell lines, but only caspase 9 in SW620 cells [54]. After 48 hours, the two caspases were fully activated. Our results here emphasize the fact that lowbush blueberry Pcys are more potent as they significantly activate both caspases 8 and 9 in SW620 (a) and SW480 (b) cell lines (Figure 6) after 24 and 48 hours. As for apple Pcys, shorter incubation times (i.e., 3 and 6 hours) resulted in caspase 8 activation only, highlighting the importance of the extrinsic apoptosis pathway.

Bottom Line: Apoptosis induction is monitored by cell surface phosphatidylserine (PS) detection.Lowbush blueberry extract triggers the strongest activity.We highlight here that berries Pcys, especially lowbush blueberry Pcys, are of putative interest for nutritional chemoprevention of colorectal cancer in view of their apoptosis induction in a human colorectal cancer cell lines.

View Article: PubMed Central - PubMed

Affiliation: Laboratoire d'Innovation Thérapeutique, UMR 7200, Faculté de Pharmacie, Université de Strasbourg, 74 route du Rhin, BP 24, 67401 Illkirch Cedex, France.

ABSTRACT

Scope: The aim of this work is to identify which proapoptotic pathway is induced in human colon cancer cell lines, in contact with proanthocyanidins extracted from various berries.

Methods and results: Proanthocyanidins (Pcys) extracted from 11 berry species are monitored for proapoptotic activities on two related human colon cancer cell lines: SW480-TRAIL-sensitive and SW620-TRAIL-resistant. Apoptosis induction is monitored by cell surface phosphatidylserine (PS) detection. Lowbush blueberry extract triggers the strongest activity. When tested on the human monocytic cell line THP-1, blueberry Pcys are less effective for PS externalisation and DNA fragmentation is absent, highlighting a specificity of apoptosis induction in gut cells. In Pcys-treated gut cell lines, caspase 8 (apoptosis extrinsic pathway) but not caspase 9 (apoptosis intrinsic pathway) is activated after 3 hours through P38 phosphorylation (90 min), emphasizing the potency of lowbush blueberry Pcys to eradicate gut TRAIL-resistant cancer cells.

Conclusion: We highlight here that berries Pcys, especially lowbush blueberry Pcys, are of putative interest for nutritional chemoprevention of colorectal cancer in view of their apoptosis induction in a human colorectal cancer cell lines.

No MeSH data available.


Related in: MedlinePlus