Targeting cisplatin-resistant human tumor cells with metabolic inhibitors (original) (raw)

Abstract

Purpose

Although cisplatin is the drug of choice in treating lung cancer patients, relapse and resistance is a common drawback to its clinical effectiveness. Based on cisplatin’s reported ability to interfere with numerous cellular components, including mitochondria, we probed alterations in metabolism in cisplatin-resistant tumor cell lines to reveal targets for overcoming this important form of resistance.

Methods

Cisplatin-resistant lung and ovarian cancer cell lines were used to evaluate the efficacy of metabolic inhibitors for selectively targeting cisplatin-resistant cells under varying oxygen conditions.

Results

Three cisplatin-resistant cancer cell lines expressed lower HKII protein when compared to the respective cisplatin-sensitive cancer cell lines from which they were derived. Under anaerobic and hypoxic conditions, treatment with the glycolytic inhibitors 2-deoxyglucose (2-DG) and 2-fluorodeoxyglucose (2-FDG) correlated with increased cytotoxicity and more pronounced decreases in lactate production in cisplatin-resistant cells, indicating a greater blockage of glycolysis. Knockdown of HKI or HKII with siRNA in the parental lung cancer cell lines led to increased 2-FDG-induced cell death under anaerobic conditions. Under normal oxygen conditions, blockage of either fatty acid oxidation or deprivation of glutamine resulted in cell death in cisplatin-resistant lung cancer cell lines.

Conclusions

Altered hexokinase levels in cisplatin-resistant cancer cell lines leads to increased sensitivity to glycolytic inhibition under anaerobic conditions, whereas under normoxic conditions, blockage of either fatty acid oxidation or deprivation of glutamine leads to cell death. These findings may be clinically applicable when considering cisplatin resistance.

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Acknowledgments

This work was supported by the National Cancer Institute Grant #CA37109 to T.J.L. The authors would also like to acknowledge Dr. Niramol Savaraj for her contribution of the SCLC and NSCLC cell lines.

Author information

Authors and Affiliations

  1. Department of Cell Biology and Anatomy, University of Miami Miller School of Medicine, P.O. Box 016960 (R124), Miami, FL, 33101, USA
    Elizabeth J. Sullivan, Metin Kurtoglu, Huaping Liu & Theodore J. Lampidis
  2. Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, PAP Building, Room 115, 1550 NW 10th Ave, Miami, FL, 33136, USA
    Randall Brenneman & Theodore J. Lampidis

Authors

  1. Elizabeth J. Sullivan
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  2. Metin Kurtoglu
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  3. Randall Brenneman
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  4. Huaping Liu
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  5. Theodore J. Lampidis
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Corresponding author

Correspondence toTheodore J. Lampidis.

Electronic supplementary material

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280_2013_2366_MOESM1_ESM.jpg

Oligomycin treatment under normoxia does not increase HK protein levels. a Immunoblot of lung cancer cell lines treated with oligomycin for 24 h. β-actin was used as a loading control. b 24-h 2-DG treatment under normoxia is not cytotoxic in either cisplatin-sensitive SCLC1 or cisplatin-resistant SR2. c 2-DG treatment under normoxia did not induce cell death in NSCLCS or NSCLCSC after 24 h. d 24-h 2-FDG treatment under normoxia is not sufficient to induce cell death in SCLC1 or SR2. e 2-FDG treatment did not induce cell death in cisplatin-sensitive NSCLCS or cisplatin-resistant NSCLCSC. (JPG 373 kb)

280_2013_2366_MOESM2_ESM.jpg

a Immunoblot probing lung cancer cell lines for HIF-1α, HKII, and HKI levels under 0.5 % O2 hypoxia. β-actin was used as a loading control. b Densitometry of HKII immunoblot for lung cancer cell lines under hypoxia. HKII is lower in cisplatin-resistant SR2 and NSCLCSC than in cisplatin-sensitive parental SCLC1 and NSCLCS, respectively. c Densitometry of HKI immunoblot for lung cancer cell lines under hypoxia. HKI expression is reduced in cisplatin-resistant cell lines compared to cisplatin-sensitive parental cell lines. (JPG 503 kb)

280_2013_2366_MOESM3_ESM.jpg

CPT-1a protein levels are not elevated in cisplatin-resistant NSCLCSC under normoxia compared with cisplatin-sensitive NSCLCS. β-actin was used as a loading control (JPG 148 kb)

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Sullivan, E.J., Kurtoglu, M., Brenneman, R. et al. Targeting cisplatin-resistant human tumor cells with metabolic inhibitors.Cancer Chemother Pharmacol 73, 417–427 (2014). https://doi.org/10.1007/s00280-013-2366-8

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