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Glucose-regulated protein 78 mediates the therapeutic efficacy of 17-DMAG in colon cancer cells

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Tumor Biology

Abstract

Glucose-regulated protein 78 (GRP78) is expressed as part of the molecular response to endoplasmic reticulum (ER) stress and mediates protein folding within the cell. GRP78 is also an important biomarker of cancer progression and the therapeutic response of patients with different cancer types. However, the role of GRP78 in the cytotoxic effect of 17-DMAG in colon cancer cells remains unclear. GRP78 expression was knocked down by small interfering RNA (siRNA). The anticancer effects of 17-DMAG were assessed by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, a flow cytometric cell-cycle analysis, and an Annexin V-propidium iodide (PI) apoptotic assay. We found that HT-29 cells expressed a lower level of GRP78 compared with DLD-1 cells. The MTT assay revealed that HT-29 cells were more sensitive to 17-DMAG treatment than DLD-1 cells. GRP78 knock down (GRP78KD) cells demonstrated an increased sensitivity to 17-DMAG treatment compared with the scrambled control cells. Based on the cell-cycle analysis and Annexin V-PI apoptotic assay, apoptosis dramatically increased in GRP78KD cells compared with scrambled control DLD-1 cells after these cells were treated with 17-DMAG. Finally, we observed a decrease in the level of Bcl-2 and an increase in the levels of Bad and Bax in GRP78KD cells treated with 17-DMAG. These results are consistent with an increased sensitivity to 17-DMAG after knock down of GRP78. The level of GRP78 expression may determine the therapeutic efficacy of 17-DMAG against colon cancer cells.

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Abbreviations

GRP78:

Glucose-regulated protein 78

ER:

Endoplasmic reticulum

17-DMAG:

17-Dimethylaminoethylamino-17-demethoxygeldanamycin

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Acknowledgments

This study was supported by grants from the National Science Council (NSC101-2314-B-038-029-MY3 and NSC101-2314-B-038-016-MY3).

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Authors and Affiliations

  1. Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan

    Yu-Jia Chang

  2. Department of Surgery, School of Medicine, College of Medicine, Taipei Medical University, 250 Wu-Xing Street, Taipei, 110, Taiwan

    Yu-Jia Chang, Chin-Sheng Hung & Po-Li Wei

  3. Division of General Surgery, Department of Surgery, Taipei Medical University Hospital, Taipei Medical University, Taipei, Taiwan

    Yu-Jia Chang, Chin-Sheng Hung & Po-Li Wei

  4. Cancer Research Center, Taipei Medical University Hospital, Taipei Medical University, Taipei, Taiwan

    Yu-Jia Chang, Chin-Sheng Hung & Po-Li Wei

  5. Division of General Surgery, Department of Surgery, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan

    Chien-Yu Huang

  6. Department of Neurosurgery, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan

    Chien-Yu Huang

  7. Department of Public Health, School of Public Health, Taipei Medical University, Taipei, Taiwan

    Hui-Hsiung Liu

  8. Graduate Institute of Cancer Biology and Drug Discovery, Taipei Medical University, Taipei, Taiwan

    Po-Li Wei

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Correspondence to Po-Li Wei.

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Chang, YJ., Huang, CY., Hung, CS. et al. Glucose-regulated protein 78 mediates the therapeutic efficacy of 17-DMAG in colon cancer cells. Tumor Biol. 36, 4367–4376 (2015). https://doi.org/10.1007/s13277-015-3076-0

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