Inhibition of glucose turnover by 3-bromopyruvate counteracts pancreatic cancer stem cell features and sensitizes cells to gemcitabine.

Abstract

According to the cancer stem cell (CSC) hypothesis, the aggressive growth and early metastasis of pancreatic ductal adenocarcinoma (PDA) is due to the activity of CSCs, which are not targeted by current therapies. Otto Warburg suggested that the growth of cancer cells is driven by a high glucose metabolism. Here, we investigated whether glycolysis inhibition targets CSCs and thus may enhance therapeutic efficacy. Four established and 3 primary PDA cell lines, non-malignant cells, and 3 patient-tumor-derived CSC-enriched spheroidal cultures were analyzed by glucose turnover measurements, MTT and ATP assays, flow cytometry of ALDH1 activity and annexin positivity, colony and spheroid formation, western blotting, electrophoretic mobility shift assay, xenotransplantation, and immunohistochemistry. The effect of siRNA-mediated inhibition of LDH-A and LDH-B was also investigated. The PDA cells exhibited a high glucose metabolism, and glucose withdrawal or LDH inhibition by siRNA prevented growth and colony formation. Treatment with the anti-glycolytic agent 3-bromopyruvate almost completely blocked cell viability, self-renewal potential, NF-κB binding activity, and stem cell-related signaling and reverted gemcitabine resistance. 3-bromopyruvate was less effective in weakly malignant PDA cells and did not affect non-malignant cells, predicting minimal side effects. 3-bromopyruvate inhibited in vivo tumor engraftment and growth on chicken eggs and mice and enhanced the efficacy of gemcitabine by influencing the expression of markers of proliferation, apoptosis, self-renewal, and metastasis. Most importantly, primary CSC-enriched spheroidal cultures were eliminated by 3-bromopyruvate. These findings propose that CSCs may be specifically dependent on a high glucose turnover and suggest 3-bromopyruvate for therapeutic intervention.

• Authors: Orkhan Isayev, Vanessa Rausch, Nathalie Bauer, Li Liu, Pei Fan, Yiyao Zhang, Jury Gladkich, Clifford C Nwaeburu, Jürgen Mattern, Martin Mollenhauer, Felix Rückert, Sebastian Zach, Uwe Haberkorn, Wolfgang Gross, Frank Schönsiegel, Alexandr V Bazhin, Ingrid Herr
• Journal: Oncotarget (Oncotarget) Vol. 5 Issue 13 Pg. 5177-89 (Jul 15 2014) ISSN: 1949-2553 [Electronic] United States
• PMID: 25015789 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antimetabolites, Antineoplastic
• Biomarkers, Tumor
• Isoenzymes
• Pyruvates
• Deoxycytidine
• bromopyruvate
• L-Lactate Dehydrogenase
• Glucose
• Gemcitabine

Topics

• Animals
• Antimetabolites, Antineoplastic (pharmacology)
• Biomarkers, Tumor (metabolism)
• Blotting, Western
• Carcinoma, Pancreatic Ductal (genetics, metabolism, prevention & control)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Chick Embryo
• Deoxycytidine (analogs & derivatives, pharmacology)
• Female
• Glucose (metabolism)
• Glycolysis (drug effects)
• Humans
• Immunohistochemistry
• Isoenzymes (genetics, metabolism)
• L-Lactate Dehydrogenase (genetics, metabolism)
• Mice, Inbred Strains
• Mice, Nude
• Neoplastic Stem Cells (drug effects, metabolism)
• Pancreatic Neoplasms (genetics, metabolism, prevention & control)
• Pyruvates (pharmacology)
• RNA Interference
• Tumor Burden (drug effects)
• Xenograft Model Antitumor Assays
• Gemcitabine