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Synergistic increase in efficacy of a combination of 2-deoxy-d-glucose and cisplatin in normoxia and hypoxia: switch from autophagy to apoptosis

  • Original Article
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Tumor Biology

Abstract

Resistance to drugs, which is aggravated by hypoxia, is a well-known feature of tumors. The combination of drug exposure and hypoxia can give rise to several survival strategies in the exposed cells. Glioblastoma multiforme (GBM) is among the most hypoxic of solid tumors, and we have used glial cells to identify a drug combination that would be synergistically effective in these cells under both normoxia and hypoxia. Cisplatin (CP) and 2-deoxy-d-glucose (2-DG), which have been used for second-line therapy and for preclinical research, are relatively ineffective as single agents. During in vitro experiments with A172 and LN229 cells, there was increased resistance to both drugs under hypoxia. However, the combination of CP and 2-DG showed a synergistic effect in reducing cell viability under both normoxia and hypoxia, with a combination index of less than 1. Increased autophagy is a distinct feature of the response to 2-DG. However, autophagic markers were reduced, and apoptotic markers were upregulated by the combination, indicating a switch over from autophagic to apoptotic pathways with reduction in endoplasmic reticulum (ER) stress. The combination also resulted in a decrease of pAKT levels. The effect of CP in the combination was replicated by the prototype AKT inhibitor LY294002, further supporting the role of AKT inhibition in the synergism. Combination of 2-DG with CP, or possibly an AKT inhibitor, can prove to be an effective rational combination for reducing chemoresistance under both normoxic and hypoxic conditions in gliomas.

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Abbreviations

GBM:

Glioblastoma multiforme

UPR:

Unfolded protein response

2-DG:

2-Deoxy-d-glucose

CP:

Cisplatin

ER:

Endoplasmic reticulum

ATP:

Adenosine triphosphate

AMPK:

AMP-activated protein kinase

TMZ:

Temozolomide

ATCC:

American Type Culture Collection

CI:

Combination index

3-MA:

3-Methyladenine

CQ:

Chloroquine

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Acknowledgments

The study has been supported by core institutional grant of the National Brain Research Centre (NBRC), India to Prof. Subrata Sinha. Fellowships provided by the Indian Council of Medical Research (ICMR) to Akansha Jalota and Mukesh Kumar are acknowledged. Prof. B. C. Das is thankful to the Department of Science and Technology (DST) for the award of J C Bose fellowship and Dr. Ajay Kumar Yadav to the Department of Biotechnology (DBT) for Ramalingaswamy fellowship. The technical support provided by Ms. Jyoti, Mr. Pappu Prasad, and Mr. Mukesh is deeply acknowledged.

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Authors and Affiliations

  1. Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, 110029, India

    Akansha Jalota, Kunzang Chosdol & Subrata Sinha

  2. Dr. B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, 110007, India

    Akansha Jalota, Bhudev C. Das & Ajay K. Yadav

  3. National Brain Research Centre, Manesar, Gurgaon, 122051, India

    Mukesh Kumar & Subrata Sinha

  4. Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Noida, 201313, India

    Bhudev C. Das

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  1. Akansha Jalota
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Correspondence to Subrata Sinha.

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Jalota, A., Kumar, M., Das, B.C. et al. Synergistic increase in efficacy of a combination of 2-deoxy-d-glucose and cisplatin in normoxia and hypoxia: switch from autophagy to apoptosis. Tumor Biol. 37, 12347–12358 (2016). https://doi.org/10.1007/s13277-016-5089-8

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