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Combined treatment of AT101 and demethoxycurcumin yields an enhanced anti-proliferative effect in human primary glioblastoma cells

  • Original Article – Cancer Research
  • Published:
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Abstract

Purpose

Glioblastoma multiforme (GBM) is a poorly curable disease due to its profound chemoresistance. Despite recent advances in surgery, radiotherapy and chemotherapy, the efficient treatment of GBMs is still a clinical challenge. Beside others, AT101, the R-(−) enantiomer of gossypol, and demethoxycurcumin (DMC), a curcumin-related demethoxy compound derived from Curcuma longa, were considered as possible alternative drugs for GBM therapy.

Methods

Using different human primary GBM cell cultures in a long-term stimulation in vitro model, the cytotoxic and anti-proliferative effects of single and combined treatment with 5 µM AT101 and 5 µM or 10 µM DMC were investigated. Furthermore, western blots on pAkt and pp44/42 as well as JC-1 staining and real-time RT-PCR were performed to understand the influence of the treatment at the molecular and gene level.

Results

Due to enhanced anti-proliferative effects, we showed that combined therapy with both drugs was superior to a single treatment with AT101 or DMC. Here, by determination of the combination index, a synergism of the combined drugs was detectable. Phosphorylation and thereby activation of the kinases p44/42 and Akt, which are involved in proliferation and survival processes, were inhibited, the mitochondrial membrane potential of the GBM cells was altered, and genes involved in dormancy-associated processes were regulated by the combined treatment strategy.

Conclusion

Combined treatment with different drugs might be an option to efficiently overcome chemoresistance of GBM cells in a long-term treatment strategy.

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Abbreviations

BSA:

Bovine serum albumin

CI:

Combination index

DMC:

Demethoxycurcumin

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

Dimethylsulfoxide

EphA5:

Ephrin type-A receptor 5

FBS:

Fetal bovine serum

GAPDH:

Glycerinaldehyde 3-phosphate dehydrogenase

GBM:

Glioblastoma multiforme

H2BK:

Histone cluster 1 H2B family member K

IGFBP5:

Insulin-like growth factor-binding protein 5

OD:

Optical density

PBS:

Phosphate-buffered saline

PC:

Primary culture

qRT-PCR:

Quantitative real-time polymerase chain reaction

TBS-T:

Tris-buffered saline with 0.1% Tween

TMZ:

Temozolomide

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Acknowledgements

We thank Fereshteh Ebrahim and Brigitte Rehmke for expert technical assistance.

Funding

This work was funded by the German Research Foundation (DFG) as part of the Research Training Group “Materials4Brain” (RTG2154; P8).

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Authors and Affiliations

  1. Department of Neurosurgery, University Medical Center Schleswig-Holstein UKSH, Campus Kiel, Arnold-Heller-Str.3, Building 41, 24105, Kiel, Germany

    Moiken Mehner, Carolin Kubelt, Vivian Adamski, Michael Synowitz & Janka Held-Feindt

  2. Department of Anatomy, University of Kiel, 24118, Kiel, Germany

    Christina Schmitt

Authors
  1. Moiken Mehner
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  2. Carolin Kubelt
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  3. Vivian Adamski
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  4. Christina Schmitt
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  5. Michael Synowitz
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  6. Janka Held-Feindt
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Contributions

JHF, CK, and CS conceived and designed the study; MM, VA, CS, and JHF performed the experiments and analyzed the data; CK and MS contributed materials and data and assisted in data analysis; JHF and MM wrote the paper; and all authors revised the manuscript.

Corresponding author

Correspondence to Janka Held-Feindt.

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Conflict of interest

Author Moiken Mehner declares that she has no conflict of interest. Author Carolin Kubelt declares that she has no conflict of interest. Author Vivian Adamski declares that she has no conflict of interest. Author Christina Schmitt declares that she has no conflict of interest. Author Michael Synowitz declares that he has no conflict of interest. Author Janka Held-Feindt declares that she has no conflict of interest.

Ethical approval

All procedures performed in studies involving human tumor samples were in accordance with the ethical standards of the institutional committee (ethics committee of the University of Kiel, Germany; file references: D471/15 and D524/17) and with the 1964 Helsinki Declaration and its later amendments. Written informed consent was obtained from all individual donors included in the study.

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Mehner, M., Kubelt, C., Adamski, V. et al. Combined treatment of AT101 and demethoxycurcumin yields an enhanced anti-proliferative effect in human primary glioblastoma cells. J Cancer Res Clin Oncol 146, 117–126 (2020). https://doi.org/10.1007/s00432-019-03107-7

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