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Targeted induction of apoptosis in glioblastoma multiforme cells by an MRP3-specific TRAIL fusion protein in vitro

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Tumor Biology

Abstract

Single-chain Fv fragments (scFvs) consist of the variable heavy-chain (VH) and variable light-chain (VL) domains, which are the smallest immunoglobulin fragments containing the whole antigen-binding site. Human soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) proves to acquire a potent pro-apoptotic activity only after selective binding to a predefined tumor cell surface antigen and has no off-target effects towards normal cells. Glioblastoma multiforme (GBM) is the most frequent and aggressive type of brain tumor and overexpresses human multidrug resistance protein 3 (MRP3). In this study, we designed a novel fusion protein, termed scFvM58-sTRAIL, in which the MRP3-specific scFv antibody M58 was genetically fused to the N-terminus of human soluble TRAIL (sTRAIL). The recombinant scFvM58-sTRAIL fusion protein, expressed in Escherichia coli, was purified by chromatography and tested for cytotoxicity. scFvM58-sTRAIL showed a significant apoptosis-inducing activity towards MRP3-positive GBM cells in vitro. The pro-apoptotic activity of scFvM58-sTRAIL towards GBM cells was strongly inhibited in the presence of the parental scFvM58 antibody, suggesting that cytotoxic activity is MRP3-restricted. In a control experiment with MRP3-negative Jurkat cells, scFvM58-sTRAIL did not induce apparent apoptosis. In addition, through target antigen-restricted binding, scFvM58-sTRAIL was capable of activating not only TRAIL-R1 but also TRAIL-R2. In conclusion, our results suggest that fusion protein scFvM58-sTRAIL with specificity for MRP3 is a highly selective therapeutic agent and may provide an alternative therapy for human GBM.

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Abbreviations

GBM:

Glioblastoma multiforme

MRP3:

Multidrug resistance protein 3

TRAIL:

Tumor necrosis factor-related apoptosis-inducing ligand

scFv:

Single-chain Fv antibody fragment

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Acknowledgments

The study was supported by grants from the Natural Science Foundation (No: 30571906) and the Key New Drug Creation of China (No: 2009ZX09102-234 and 2009ZX09103-689).

Conflicts of interest

None.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Second Military Medical University, Xiangyin Road No. 800, Shanghai, 200433, China

    Liang-Hua Wang, Cui-Ting Lv, Yuan Le & Bing-Hua Jiao

  2. Department of Neurology, Second Affiliated Hospital of Dalian Medical University, Dalian, 116023, China

    Chang-Wei Ni, Yong-Zhong Lin & Lin Yin

  3. Department of Neurology, First Affiliated Hospital of Dalian Medical University, Dalian, 116011, China

    Chang-Bin Jiang

  4. Epilepsy Center of Liaoning Province, Second Affiliated Hospital of Dalian Medical University, Dalian, 116023, China

    Yue Lang

  5. College of Pharmacy, Dalian Medical University, Dalian, 116085, China

    Chen-Yang Zhao

  6. Department of Neurosurgery, Second Affiliated Hospital of Dalian Medical University, Zhongshan Road No. 467, Dalian, 116023, China

    Kang Yang & Jian Yin

Authors
  1. Liang-Hua Wang
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  2. Chang-Wei Ni
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  7. Yuan Le
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  8. Yue Lang
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  9. Chen-Yang Zhao
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  10. Kang Yang
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  11. Bing-Hua Jiao
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  12. Jian Yin
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Corresponding authors

Correspondence to Bing-Hua Jiao or Jian Yin.

Additional information

Liang-Hua Wang and Chang-Wei Ni contributed equally to this article.

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Wang, LH., Ni, CW., Lin, YZ. et al. Targeted induction of apoptosis in glioblastoma multiforme cells by an MRP3-specific TRAIL fusion protein in vitro. Tumor Biol. 35, 1157–1168 (2014). https://doi.org/10.1007/s13277-013-1155-7

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  • DOI: https://doi.org/10.1007/s13277-013-1155-7

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