PMC

Tumor-cell induced aggregation, thrombus formation and activation are decreased by 3-BP. (A) Platelets were pre-incubated with CRC cells (1.5 × 10⁴ cells) and 3-BP (100 μM) for 5 min, and subsequently stimulated using collagen (2 μg/mL). Aggregation was measured for 10 min. Platelets stimulated with collagen only served as controls for the experiment. Each data point (special shapes) corresponds to an individual experiment and indicates the data for platelets marked with: Circle—only platelets, Square—with HCT116, Up-triangle—with HCT116 and 3BP, Down-triangle—with HT29, Diamond—with HT29 and 3BP, Hollow circle—with Caco-2, Hollow Square—with Caco-2 and 3BP. (B) Co-cultures of tumor cells and platelets were visualized by microscopy. Platelet aggregation as assessed by microscopy is indicated with arrows and quantified (C). (D) Platelets from co-culture with CRC cells (HCT116 and Caco-2) were lysed and loaded for western blot analysis of platelet activation markers and phosphatases. β-actin served as loading control. Densitometric values, corrected for loading control, are indicated. A representative blot of at least 2 independent experiments is shown. (E–G) Platelets co-cultured with HT29 (E), HCT116 (F) or Caco-2 (G) cells in the absence or presence of 3-BP were subsequently subjected to FACS analysis to determine cell surface expression of vWF-receptor (CD41-FITC), Integrinβ3 (CD42-PE) and P-Selectin (CD62-APC). Each data point (special shapes) corresponds to an individual experiment and indicates the data as: Circle—Integrinβ3 without 3BP, Square—Integrinβ3 with 3BP, Up-triangle—vWF-receptor without 3BP, Down-triangle—vWF-receptor with 3BP, Diamond—P-Selectin without 3BP, Hollow circle—P-Selectin with 3BP.

3BP structure and delivery.
Notes: (A) 3BP is similar in structure to pyruvate (precursor of Krebs cycle), lactate (Warburg effect) and acetate (precursor of lipogenesis). 3BP acts as an antimetabolite to these vital metabolic substrates for cancer cells. (B) Liposomal formulations of 3BP may improve delivery, metabolism, targeting, monitoring and potentiate 3BP-induced anticancer effects.
Abbreviations: 3BP, 3-Bromopyruvate; EGF, epidermal growth factor; PEG, polyethylene glycol.

A, treatment doses given to the patient. B, 3BP treatment caused a moderate decrease in serum lactate dehydrogenase (LDH) level, a metabolic predictor of cancer cell energy. Serum LDH level decreased maximally on combining paracetamol and 3BP.

HCT116 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)).