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The effect of 3-bromopyruvate on human colorectal cancer cells is dependent on glucose concentration but not hexokinase II expression.

Ho N, Morrison J, Silva A, Coomber BL - Biosci. Rep. (2016)

Bottom Line: High HKII-expressing cell lines were more sensitive to 3BP than low HKII-expressing cells. 3BP-induced rapid Akt phosphorylation at site Thr-308 and cell death via both apoptotic and necrotic mechanisms.Cells grown under lower glucose concentrations showed greater resistance towards 3BP.Cells with HKII knockdown showed no changes in 3BP sensitivity, suggesting the effects of 3BP are independent of HKII expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada, N1G 2W1.

No MeSH data available.


Related in: MedlinePlus

Type of cell death induced by 3BP is dose-dependentCaspase-3 cleavage and DNA fragmentation was assessed to further elucidate the mechanisms of 3BP-induced cell death. Caspase-3 cleavage was examined through western blot analysis (N=3) (A). An image of a longer exposed membrane to better detect cleaved caspase-3 is shown below each full caspase-3 blot. DNA fragmentation was examined through gel electrophoresis in SW480 cells following 48 h 3BP treatment using 0.8% (B) and 2% (C) gels (N=2). Etoposide (E; 50 μM) was used as a positive control for caspase-3 cleavage and DNA fragmentation; note nucleosome laddering in etoposide treated cells in (C) indicative of apoptosis.
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Figure 3: Type of cell death induced by 3BP is dose-dependentCaspase-3 cleavage and DNA fragmentation was assessed to further elucidate the mechanisms of 3BP-induced cell death. Caspase-3 cleavage was examined through western blot analysis (N=3) (A). An image of a longer exposed membrane to better detect cleaved caspase-3 is shown below each full caspase-3 blot. DNA fragmentation was examined through gel electrophoresis in SW480 cells following 48 h 3BP treatment using 0.8% (B) and 2% (C) gels (N=2). Etoposide (E; 50 μM) was used as a positive control for caspase-3 cleavage and DNA fragmentation; note nucleosome laddering in etoposide treated cells in (C) indicative of apoptosis.

Mentions: SW480 cells showed considerable caspase-3 cleavage even under control conditions. HCT116 cells were most resistant to etoposide-induced caspase-3 cleavage compared to CaCo2, SW480 and DLD-1 cells. Following 3BP exposure, no cleaved caspase-3 was observed in HCT116 or CaCo2 cells whereas limited caspase-3 cleavage was observed in SW480 and DLD-1 cells (Figure 3A). However, cells exposed to 50 μM 3BP showed overall reduced cellular protein as evidenced by reduced full-length caspase-3 but no detectable cleaved product, and reduced α-tubulin expression. This observation, suggestive of cellular necrosis, was verified using DNA fragmentation analysis, where 50 μM 3BP treatment of SW480 cells led to DNA smearing on the agarose gel, and no obvious nucleosome laddering expected with apoptotic cell death (Figures 3B and 3C).


The effect of 3-bromopyruvate on human colorectal cancer cells is dependent on glucose concentration but not hexokinase II expression.

Ho N, Morrison J, Silva A, Coomber BL - Biosci. Rep. (2016)

Type of cell death induced by 3BP is dose-dependentCaspase-3 cleavage and DNA fragmentation was assessed to further elucidate the mechanisms of 3BP-induced cell death. Caspase-3 cleavage was examined through western blot analysis (N=3) (A). An image of a longer exposed membrane to better detect cleaved caspase-3 is shown below each full caspase-3 blot. DNA fragmentation was examined through gel electrophoresis in SW480 cells following 48 h 3BP treatment using 0.8% (B) and 2% (C) gels (N=2). Etoposide (E; 50 μM) was used as a positive control for caspase-3 cleavage and DNA fragmentation; note nucleosome laddering in etoposide treated cells in (C) indicative of apoptosis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4759612&req=5

Figure 3: Type of cell death induced by 3BP is dose-dependentCaspase-3 cleavage and DNA fragmentation was assessed to further elucidate the mechanisms of 3BP-induced cell death. Caspase-3 cleavage was examined through western blot analysis (N=3) (A). An image of a longer exposed membrane to better detect cleaved caspase-3 is shown below each full caspase-3 blot. DNA fragmentation was examined through gel electrophoresis in SW480 cells following 48 h 3BP treatment using 0.8% (B) and 2% (C) gels (N=2). Etoposide (E; 50 μM) was used as a positive control for caspase-3 cleavage and DNA fragmentation; note nucleosome laddering in etoposide treated cells in (C) indicative of apoptosis.
Mentions: SW480 cells showed considerable caspase-3 cleavage even under control conditions. HCT116 cells were most resistant to etoposide-induced caspase-3 cleavage compared to CaCo2, SW480 and DLD-1 cells. Following 3BP exposure, no cleaved caspase-3 was observed in HCT116 or CaCo2 cells whereas limited caspase-3 cleavage was observed in SW480 and DLD-1 cells (Figure 3A). However, cells exposed to 50 μM 3BP showed overall reduced cellular protein as evidenced by reduced full-length caspase-3 but no detectable cleaved product, and reduced α-tubulin expression. This observation, suggestive of cellular necrosis, was verified using DNA fragmentation analysis, where 50 μM 3BP treatment of SW480 cells led to DNA smearing on the agarose gel, and no obvious nucleosome laddering expected with apoptotic cell death (Figures 3B and 3C).

Bottom Line: High HKII-expressing cell lines were more sensitive to 3BP than low HKII-expressing cells. 3BP-induced rapid Akt phosphorylation at site Thr-308 and cell death via both apoptotic and necrotic mechanisms.Cells grown under lower glucose concentrations showed greater resistance towards 3BP.Cells with HKII knockdown showed no changes in 3BP sensitivity, suggesting the effects of 3BP are independent of HKII expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada, N1G 2W1.

No MeSH data available.


Related in: MedlinePlus