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Apoptotic effect of a novel kefir product, PFT, on multidrug-resistant myeloid leukemia cells via a hole-piercing mechanism.

Ghoneum M, Gimzewski J - Int. J. Oncol. (2014)

Bottom Line: We examined the apoptotic effect of a novel Probiotics Fermentation Technology (PFT) kefir grain product; PFT is a natural mixture composed primarily of Lactobacillus kefiri P-IF, a specific strain of L. kefiri with unique growth characteristics.The apoptotic effect of PFT was assessed through examination of percent apoptosis, caspase 3 activation, Bcl-2 expression levels and changes in mitochondrial membrane potential (MMP).In addition, PFT showed a unique characteristic of piercing holes in HL60/AR cells, as indicated by AFM studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Charles R. Drew University of Medicine and Science, Los Angeles, CA 90059, USA.

ABSTRACT
We examined the apoptotic effect of a novel Probiotics Fermentation Technology (PFT) kefir grain product; PFT is a natural mixture composed primarily of Lactobacillus kefiri P-IF, a specific strain of L. kefiri with unique growth characteristics. The aim of this study was to examine the apoptotic effect of PFT on human multidrug-resistant (MDR) myeloid leukemia (HL60/AR) cells in vitro and explore the mechanistic approach underlying its effect. HL60/AR cells were cultured with PFT (0.6-5.0 mg/ml) for 3 days. The apoptotic effect of PFT was assessed through examination of percent apoptosis, caspase 3 activation, Bcl-2 expression levels and changes in mitochondrial membrane potential (MMP). PFT induced apoptosis in HL60/AR cells in a dose-dependent manner which was maximal at 67.5% for 5 mg/ml. Induction of apoptosis was associated with activation of caspase 3, decreased expression of Bcl-2 and decreased polarization of MMP. In addition, PFT showed a unique characteristic of piercing holes in HL60/AR cells, as indicated by AFM studies. This hole induction may be responsible for the apoptotic effect on cancer cells. These results suggest that PFT may act as a potential therapy for the treatment of MDR leukemia.

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PFT induces apoptosis through activation of caspase 3. The activation of caspase 3 was determined in cancer cells post-culture with PFT at different concentrations (0, 0.6, 1.25 and 2.5 mg/ml). The lines represent cancer cells in the absence of PFT (control, purple) and cancer cells treated with PFT at concentration of 0.6 mg/ml (green), 1.25 mg/ml (pink), or 2.5 mg/ml (blue). The activation of caspase 3 in cells was detected using carboxyfluorescein-labeled cell permeable peptide substrate and was analyzed using flow cytometry.
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f3-ijo-44-03-0830: PFT induces apoptosis through activation of caspase 3. The activation of caspase 3 was determined in cancer cells post-culture with PFT at different concentrations (0, 0.6, 1.25 and 2.5 mg/ml). The lines represent cancer cells in the absence of PFT (control, purple) and cancer cells treated with PFT at concentration of 0.6 mg/ml (green), 1.25 mg/ml (pink), or 2.5 mg/ml (blue). The activation of caspase 3 in cells was detected using carboxyfluorescein-labeled cell permeable peptide substrate and was analyzed using flow cytometry.

Mentions: To verify activation of the apoptotic pathway, the level of caspase 3 activation was investigated by treating HL60/AR cells with PFT at varying concentrations for 24 h and analyzed using flow cytometry. Data depicted in Fig. 3 show that the proportion of cancer cells with increased active caspase 3 was higher in PFT-treated cells than in control untreated cells. This would suggest that pre-exposure of HL60/AR cells to PFT led to increased activation of the executioner caspase 3.


Apoptotic effect of a novel kefir product, PFT, on multidrug-resistant myeloid leukemia cells via a hole-piercing mechanism.

Ghoneum M, Gimzewski J - Int. J. Oncol. (2014)

PFT induces apoptosis through activation of caspase 3. The activation of caspase 3 was determined in cancer cells post-culture with PFT at different concentrations (0, 0.6, 1.25 and 2.5 mg/ml). The lines represent cancer cells in the absence of PFT (control, purple) and cancer cells treated with PFT at concentration of 0.6 mg/ml (green), 1.25 mg/ml (pink), or 2.5 mg/ml (blue). The activation of caspase 3 in cells was detected using carboxyfluorescein-labeled cell permeable peptide substrate and was analyzed using flow cytometry.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928472&req=5

f3-ijo-44-03-0830: PFT induces apoptosis through activation of caspase 3. The activation of caspase 3 was determined in cancer cells post-culture with PFT at different concentrations (0, 0.6, 1.25 and 2.5 mg/ml). The lines represent cancer cells in the absence of PFT (control, purple) and cancer cells treated with PFT at concentration of 0.6 mg/ml (green), 1.25 mg/ml (pink), or 2.5 mg/ml (blue). The activation of caspase 3 in cells was detected using carboxyfluorescein-labeled cell permeable peptide substrate and was analyzed using flow cytometry.
Mentions: To verify activation of the apoptotic pathway, the level of caspase 3 activation was investigated by treating HL60/AR cells with PFT at varying concentrations for 24 h and analyzed using flow cytometry. Data depicted in Fig. 3 show that the proportion of cancer cells with increased active caspase 3 was higher in PFT-treated cells than in control untreated cells. This would suggest that pre-exposure of HL60/AR cells to PFT led to increased activation of the executioner caspase 3.

Bottom Line: We examined the apoptotic effect of a novel Probiotics Fermentation Technology (PFT) kefir grain product; PFT is a natural mixture composed primarily of Lactobacillus kefiri P-IF, a specific strain of L. kefiri with unique growth characteristics.The apoptotic effect of PFT was assessed through examination of percent apoptosis, caspase 3 activation, Bcl-2 expression levels and changes in mitochondrial membrane potential (MMP).In addition, PFT showed a unique characteristic of piercing holes in HL60/AR cells, as indicated by AFM studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Charles R. Drew University of Medicine and Science, Los Angeles, CA 90059, USA.

ABSTRACT
We examined the apoptotic effect of a novel Probiotics Fermentation Technology (PFT) kefir grain product; PFT is a natural mixture composed primarily of Lactobacillus kefiri P-IF, a specific strain of L. kefiri with unique growth characteristics. The aim of this study was to examine the apoptotic effect of PFT on human multidrug-resistant (MDR) myeloid leukemia (HL60/AR) cells in vitro and explore the mechanistic approach underlying its effect. HL60/AR cells were cultured with PFT (0.6-5.0 mg/ml) for 3 days. The apoptotic effect of PFT was assessed through examination of percent apoptosis, caspase 3 activation, Bcl-2 expression levels and changes in mitochondrial membrane potential (MMP). PFT induced apoptosis in HL60/AR cells in a dose-dependent manner which was maximal at 67.5% for 5 mg/ml. Induction of apoptosis was associated with activation of caspase 3, decreased expression of Bcl-2 and decreased polarization of MMP. In addition, PFT showed a unique characteristic of piercing holes in HL60/AR cells, as indicated by AFM studies. This hole induction may be responsible for the apoptotic effect on cancer cells. These results suggest that PFT may act as a potential therapy for the treatment of MDR leukemia.

Show MeSH
Related in: MedlinePlus