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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

Glucose availability alters 3BP sensitivity in CRC cellsHCT116 and DLD-1 cells were treated with 3BP under different media glucose concentrations. HKII expression was assessed via western blot analysis. Representative blots from three biological replicates (A) and graphical outputs for densitometric analysis of HKII expression normalized to α-tubulin from three biological replicates ± S.E.M. are shown (B). Representative blots from three biological replicates of putative 3BP targets SDH, GAPDH and PGK1 show no effects of altered glucose concentrations (C). Growth curves for CRC cells grown under different media glucose concentrations in the presence or absence of 3BP over 72 h (D). Dose-effects of 3BP on cell growth following 72 h treatment in CRC cells under different media glucose concentrations (E) (*P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 compared with control (0 μM 3BP)).
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Figure 5: Glucose availability alters 3BP sensitivity in CRC cellsHCT116 and DLD-1 cells were treated with 3BP under different media glucose concentrations. HKII expression was assessed via western blot analysis. Representative blots from three biological replicates (A) and graphical outputs for densitometric analysis of HKII expression normalized to α-tubulin from three biological replicates ± S.E.M. are shown (B). Representative blots from three biological replicates of putative 3BP targets SDH, GAPDH and PGK1 show no effects of altered glucose concentrations (C). Growth curves for CRC cells grown under different media glucose concentrations in the presence or absence of 3BP over 72 h (D). Dose-effects of 3BP on cell growth following 72 h treatment in CRC cells under different media glucose concentrations (E) (*P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 compared with control (0 μM 3BP)).

Mentions: HCT116 and DLD-1 cells were adapted to and maintained in DMEM with varying glucose concentrations. Cells were maintained in DMEM containing 25, 5.5 and 1 mM glucose for subsequent experimentation, denoted as ‘high’, ‘normal’ and ‘low’ glucose conditions, respectively. Western blot analysis showed that HKII levels decreased with media glucose levels in both cell lines examined (Figures 5A and 5B). Differences in HKII expression between cells grown under ‘high’ compared with ‘normal’ glucose were more profound in DLD-1 cells. Changes in glucose concentrations did not alter expression of succinate dehydrogenase (SDH), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), nor phosphoglycerate kinase (PGK1) enzymes (Figure 5C). Both CRC cell lines proliferated exponentially under ‘high’ and ‘normal’ glucose concentrations, but growth was limited in cells cultured in ‘low’ glucose over 72 h (Figure 5E). Cells grown in ‘normal’ glucose showed an increase in 3BP resistance compared with cells grown in ‘high’ glucose. Significant differences in cell growth were observed between ‘high’ and ‘normal’ glucose for HCT116 cells treated with 30 and 40 μM 3BP and DLD-1 cells treated with 20 and 30 μM 3BP (Figure 5F). This pattern is consistent with the finding that DLD-1 cells are more sensitive to 3BP than HCT116 cells.


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)

Glucose availability alters 3BP sensitivity in CRC cellsHCT116 and DLD-1 cells were treated with 3BP under different media glucose concentrations. HKII expression was assessed via western blot analysis. Representative blots from three biological replicates (A) and graphical outputs for densitometric analysis of HKII expression normalized to α-tubulin from three biological replicates ± S.E.M. are shown (B). Representative blots from three biological replicates of putative 3BP targets SDH, GAPDH and PGK1 show no effects of altered glucose concentrations (C). Growth curves for CRC cells grown under different media glucose concentrations in the presence or absence of 3BP over 72 h (D). Dose-effects of 3BP on cell growth following 72 h treatment in CRC cells under different media glucose concentrations (E) (*P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 compared with control (0 μM 3BP)).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Glucose availability alters 3BP sensitivity in CRC cellsHCT116 and DLD-1 cells were treated with 3BP under different media glucose concentrations. HKII expression was assessed via western blot analysis. Representative blots from three biological replicates (A) and graphical outputs for densitometric analysis of HKII expression normalized to α-tubulin from three biological replicates ± S.E.M. are shown (B). Representative blots from three biological replicates of putative 3BP targets SDH, GAPDH and PGK1 show no effects of altered glucose concentrations (C). Growth curves for CRC cells grown under different media glucose concentrations in the presence or absence of 3BP over 72 h (D). Dose-effects of 3BP on cell growth following 72 h treatment in CRC cells under different media glucose concentrations (E) (*P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 compared with control (0 μM 3BP)).
Mentions: HCT116 and DLD-1 cells were adapted to and maintained in DMEM with varying glucose concentrations. Cells were maintained in DMEM containing 25, 5.5 and 1 mM glucose for subsequent experimentation, denoted as ‘high’, ‘normal’ and ‘low’ glucose conditions, respectively. Western blot analysis showed that HKII levels decreased with media glucose levels in both cell lines examined (Figures 5A and 5B). Differences in HKII expression between cells grown under ‘high’ compared with ‘normal’ glucose were more profound in DLD-1 cells. Changes in glucose concentrations did not alter expression of succinate dehydrogenase (SDH), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), nor phosphoglycerate kinase (PGK1) enzymes (Figure 5C). Both CRC cell lines proliferated exponentially under ‘high’ and ‘normal’ glucose concentrations, but growth was limited in cells cultured in ‘low’ glucose over 72 h (Figure 5E). Cells grown in ‘normal’ glucose showed an increase in 3BP resistance compared with cells grown in ‘high’ glucose. Significant differences in cell growth were observed between ‘high’ and ‘normal’ glucose for HCT116 cells treated with 30 and 40 μM 3BP and DLD-1 cells treated with 20 and 30 μM 3BP (Figure 5F). This pattern is consistent with the finding that DLD-1 cells are more sensitive to 3BP than HCT116 cells.

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