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Bilberry extract (Antho 50) selectively induces redox-sensitive caspase 3-related apoptosis in chronic lymphocytic leukemia cells by targeting the Bcl-2/Bad pathway.

Alhosin M, León-González AJ, Dandache I, Lelay A, Rashid SK, Kevers C, Pincemail J, Fornecker LM, Mauvieux L, Herbrecht R, Schini-Kerth VB - Sci Rep (2015)

Bottom Line: Among the main phenolic compounds of the bilberry extract, delphinidin-3-O-glucoside and delphinidin-3-O-rutinoside induced a pro-apoptotic effect.Antho 50-induced apoptosis is associated with activation of caspase 3, down-regulation of UHRF1, a rapid dephosphorylation of Akt and Bad, and down-regulation of Bcl-2.This activity of Antho 50 involves the glucoside and rutinoside derivatives of delphinidin.

View Article: PubMed Central - PubMed

Affiliation: CNRS UMR 7213 Laboratoire de Biophotonique et Pharmacologie, Université de Strasbourg, Faculté de Pharmacie, 74, route du Rhin, 67401 Illkirch, France.

ABSTRACT
Defect in apoptosis has been implicated as a major cause of resistance to chemotherapy observed in B cell chronic lymphocytic leukaemia (B CLL). This study evaluated the pro-apoptotic effect of an anthocyanin-rich dietary bilberry extract (Antho 50) on B CLL cells from 30 patients and on peripheral blood mononuclear cells (PBMCs) from healthy subjects, and determined the underlying mechanism. Antho 50 induced concentration- and time-dependent pro-apoptotic effects in B CLL cells but little or no effect in PBMCs. Among the main phenolic compounds of the bilberry extract, delphinidin-3-O-glucoside and delphinidin-3-O-rutinoside induced a pro-apoptotic effect. Antho 50-induced apoptosis is associated with activation of caspase 3, down-regulation of UHRF1, a rapid dephosphorylation of Akt and Bad, and down-regulation of Bcl-2. Antho 50 significantly induced PEG-catalase-sensitive formation of reactive oxygen species in B CLL cells. PEG-catalase prevented the Antho 50-induced induction of apoptosis and related signaling. The present findings indicate that Antho 50 exhibits strong pro-apoptotic activity through redox-sensitive caspase 3 activation-related mechanism in B CLL cells involving dysregulation of the Bad/Bcl-2 pathway. This activity of Antho 50 involves the glucoside and rutinoside derivatives of delphinidin. They further suggest that Antho 50 has chemotherapeutic potential by targeting selectively B CLL cells.

No MeSH data available.


Related in: MedlinePlus

Antho 50 reduces cell viability and induces selectively a concentration- and time-dependent apoptosis in B CLL cells.Cells were exposed to increasing concentrations of Antho 50 for 24 h or for 75 μg/mL for the indicated times. Apoptosis in B CLL cells (A, B) and in PBMCs (D) was assessed by flow cytometry using the annexin V-FITC/PI apoptosis assay. Cell viability rate (C) was assessed by cell counting using the trypan blue dye exclusion assay. Equal quantity of genomic DNA was analyzed on a 1% agarose gel. DNA was stained with ethidium bromide and then visualized under UV light (E). The control (Ctr) represents untreated cells harvested at the latest time point. The data are representative of cells from nine CLL patients and four healthy subjects for apoptosis, six CLL patients for cell viability and two for DNA fragmentation. Values are shown as means ± S.E.M. (n = 3); *, P < 0.05, **, P < 0.01, ***, P < 0.001 versus respective control.
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f1: Antho 50 reduces cell viability and induces selectively a concentration- and time-dependent apoptosis in B CLL cells.Cells were exposed to increasing concentrations of Antho 50 for 24 h or for 75 μg/mL for the indicated times. Apoptosis in B CLL cells (A, B) and in PBMCs (D) was assessed by flow cytometry using the annexin V-FITC/PI apoptosis assay. Cell viability rate (C) was assessed by cell counting using the trypan blue dye exclusion assay. Equal quantity of genomic DNA was analyzed on a 1% agarose gel. DNA was stained with ethidium bromide and then visualized under UV light (E). The control (Ctr) represents untreated cells harvested at the latest time point. The data are representative of cells from nine CLL patients and four healthy subjects for apoptosis, six CLL patients for cell viability and two for DNA fragmentation. Values are shown as means ± S.E.M. (n = 3); *, P < 0.05, **, P < 0.01, ***, P < 0.001 versus respective control.

Mentions: To determine whether Antho 50 induces apoptosis in CLL cells, the detection of phosphatidylserine externalization by flow cytometry using annexin V FITC/PI assay kit was performed. As indicated in Fig. 1A, a concentration-dependent increase in annexin V positive cells was observed in Antho 50-treated cells for 24 h and this effect reached significance at concentrations greater than 25 μg/mL of Antho 50. The percentage of annexin V positive cells reached approximately 75% at 75 μg/mL. Incubation of cells with 75 μg/mL of Antho 50 induced a time-dependent increase in annexin V positive cells with a significant effect observed already at 1 h (Fig. 1B) and which was associated with a reduction in cell viability (Fig. 1C). To determine the selectivity of Antho 50, PBMCs from five healthy adult donors were incubated with Antho 50 for 24 h (Fig. 1D). Although Antho 50 at a concentration of 25 μg/mL significantly induced apoptosis in CLL cells by about 50% (Fig. 1A), no such effect was observed in PBMCs (Fig. 1D). However, increasing the concentration of Antho 50 to 75 μg/mL induced a slight but significant apoptosis in PBMCs by about 36% (Fig. 1D). These data indicate that Antho 50 is targeting predominantly neoplastic B cells relative to PBMCs.


Bilberry extract (Antho 50) selectively induces redox-sensitive caspase 3-related apoptosis in chronic lymphocytic leukemia cells by targeting the Bcl-2/Bad pathway.

Alhosin M, León-González AJ, Dandache I, Lelay A, Rashid SK, Kevers C, Pincemail J, Fornecker LM, Mauvieux L, Herbrecht R, Schini-Kerth VB - Sci Rep (2015)

Antho 50 reduces cell viability and induces selectively a concentration- and time-dependent apoptosis in B CLL cells.Cells were exposed to increasing concentrations of Antho 50 for 24 h or for 75 μg/mL for the indicated times. Apoptosis in B CLL cells (A, B) and in PBMCs (D) was assessed by flow cytometry using the annexin V-FITC/PI apoptosis assay. Cell viability rate (C) was assessed by cell counting using the trypan blue dye exclusion assay. Equal quantity of genomic DNA was analyzed on a 1% agarose gel. DNA was stained with ethidium bromide and then visualized under UV light (E). The control (Ctr) represents untreated cells harvested at the latest time point. The data are representative of cells from nine CLL patients and four healthy subjects for apoptosis, six CLL patients for cell viability and two for DNA fragmentation. Values are shown as means ± S.E.M. (n = 3); *, P < 0.05, **, P < 0.01, ***, P < 0.001 versus respective control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4355738&req=5

f1: Antho 50 reduces cell viability and induces selectively a concentration- and time-dependent apoptosis in B CLL cells.Cells were exposed to increasing concentrations of Antho 50 for 24 h or for 75 μg/mL for the indicated times. Apoptosis in B CLL cells (A, B) and in PBMCs (D) was assessed by flow cytometry using the annexin V-FITC/PI apoptosis assay. Cell viability rate (C) was assessed by cell counting using the trypan blue dye exclusion assay. Equal quantity of genomic DNA was analyzed on a 1% agarose gel. DNA was stained with ethidium bromide and then visualized under UV light (E). The control (Ctr) represents untreated cells harvested at the latest time point. The data are representative of cells from nine CLL patients and four healthy subjects for apoptosis, six CLL patients for cell viability and two for DNA fragmentation. Values are shown as means ± S.E.M. (n = 3); *, P < 0.05, **, P < 0.01, ***, P < 0.001 versus respective control.
Mentions: To determine whether Antho 50 induces apoptosis in CLL cells, the detection of phosphatidylserine externalization by flow cytometry using annexin V FITC/PI assay kit was performed. As indicated in Fig. 1A, a concentration-dependent increase in annexin V positive cells was observed in Antho 50-treated cells for 24 h and this effect reached significance at concentrations greater than 25 μg/mL of Antho 50. The percentage of annexin V positive cells reached approximately 75% at 75 μg/mL. Incubation of cells with 75 μg/mL of Antho 50 induced a time-dependent increase in annexin V positive cells with a significant effect observed already at 1 h (Fig. 1B) and which was associated with a reduction in cell viability (Fig. 1C). To determine the selectivity of Antho 50, PBMCs from five healthy adult donors were incubated with Antho 50 for 24 h (Fig. 1D). Although Antho 50 at a concentration of 25 μg/mL significantly induced apoptosis in CLL cells by about 50% (Fig. 1A), no such effect was observed in PBMCs (Fig. 1D). However, increasing the concentration of Antho 50 to 75 μg/mL induced a slight but significant apoptosis in PBMCs by about 36% (Fig. 1D). These data indicate that Antho 50 is targeting predominantly neoplastic B cells relative to PBMCs.

Bottom Line: Among the main phenolic compounds of the bilberry extract, delphinidin-3-O-glucoside and delphinidin-3-O-rutinoside induced a pro-apoptotic effect.Antho 50-induced apoptosis is associated with activation of caspase 3, down-regulation of UHRF1, a rapid dephosphorylation of Akt and Bad, and down-regulation of Bcl-2.This activity of Antho 50 involves the glucoside and rutinoside derivatives of delphinidin.

View Article: PubMed Central - PubMed

Affiliation: CNRS UMR 7213 Laboratoire de Biophotonique et Pharmacologie, Université de Strasbourg, Faculté de Pharmacie, 74, route du Rhin, 67401 Illkirch, France.

ABSTRACT
Defect in apoptosis has been implicated as a major cause of resistance to chemotherapy observed in B cell chronic lymphocytic leukaemia (B CLL). This study evaluated the pro-apoptotic effect of an anthocyanin-rich dietary bilberry extract (Antho 50) on B CLL cells from 30 patients and on peripheral blood mononuclear cells (PBMCs) from healthy subjects, and determined the underlying mechanism. Antho 50 induced concentration- and time-dependent pro-apoptotic effects in B CLL cells but little or no effect in PBMCs. Among the main phenolic compounds of the bilberry extract, delphinidin-3-O-glucoside and delphinidin-3-O-rutinoside induced a pro-apoptotic effect. Antho 50-induced apoptosis is associated with activation of caspase 3, down-regulation of UHRF1, a rapid dephosphorylation of Akt and Bad, and down-regulation of Bcl-2. Antho 50 significantly induced PEG-catalase-sensitive formation of reactive oxygen species in B CLL cells. PEG-catalase prevented the Antho 50-induced induction of apoptosis and related signaling. The present findings indicate that Antho 50 exhibits strong pro-apoptotic activity through redox-sensitive caspase 3 activation-related mechanism in B CLL cells involving dysregulation of the Bad/Bcl-2 pathway. This activity of Antho 50 involves the glucoside and rutinoside derivatives of delphinidin. They further suggest that Antho 50 has chemotherapeutic potential by targeting selectively B CLL cells.

No MeSH data available.


Related in: MedlinePlus