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Sulforaphane potentiates oxaliplatin-induced cell growth inhibition in colorectal cancer cells via induction of different modes of cell death

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Abstract

The objective of this study was to investigate, whether the plant-derived isothiocyanate Sulforaphane (SFN) enhances the antitumor activities of the chemotherapeutic agent oxaliplatin (Ox) in a cell culture model of colorectal cancer. Caco-2 cells were cultured under standard conditions and treated with increasing concentrations of SFN [1–20 μM] and/or Ox [100 nM–10 μM]. For co-incubation, cells were pre-treated with SFN for 24 h. Cell growth was determined by BrdU incorporation. Drug interactions were assessed using the combination-index method (CI) (Cl < 1 indicates synergism). Apoptotic events were characterized by different ELISA techniques. Protein levels were examined by Western blot analysis. Annexin V- and propidium iodide (PI) staining followed by FACS analysis was used to differentiate between apoptotic and necrotic events. SFN and Ox alone inhibited cell growth of Caco-2 cells in a dose-dependent manner, an effect, which could be synergistically enhanced, when cells were incubated with the combination of both agents. Co-treated cells further displayed distinctive morphological changes that occurred during the apoptotic process, such as cell surface exposure of phosphatidylserine, membrane blebbing as well as the occurence of cytoplasmic histone-associated DNA fragments. Further observations thereby pointed toward simultaneous activation of both extrinsic and intrinsic apoptotic pathways. With increasing concentrations and treatment duration, a shift from apoptotic to necrotic cell death could be observed. In conclusion, the data suggest that the isothiocyanate SFN sensitizes colon cancer cells to Ox-induced cell growth inhibition via induction of different modes of cell death.

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Abbreviations

CRC:

Colorectal cancer

SFN:

Sulforaphane

Ox:

Oxaliplatin

5-FU:

5-Fluorouracil

CI:

Combination index

IC50 :

Half maximal inhibitory concentration

FCS:

Fetal calf serum

DMEM:

Dulbecco’s modified Eagle’s medium

EDTA:

Ethylendiaminetetraacetic acid

DMSO:

Dimethylsulfoxid

BrdU:

Bromodeoxyuridine

TRAIL:

TNF-related apoptosis-inducing ligand

PARP:

Poly [ADP-ribose] polymerase

PI:

Propidium Iodide

FITC:

Fluorescein Isothiocyanate

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Acknowledgments

This work was supported by a graduate scholarship grant from the DFG to Bettina M. Kaminski. Bettina M. Kaminski is a member of the Frankfurt International Research Graduate School for Translational Biomedicine (FIRST), Frankfurt am Main.

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Authors and Affiliations

  1. Institute of Pharmaceutical Chemistry, Biozentrum, Goethe University, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany

    Bettina M. Kaminski, Dieter Steinhilber, Jürgen Stein & Sandra Ulrich

  2. Department of Internal Medicine, Katharina Kasper Hospital, Frankfurt am Main, Germany

    Jürgen Stein

  3. Institute of Biochemistry I/ZAFES, Goethe University, Frankfurt am Main, Germany

    Andreas Weigert & Bernhard Brüne

  4. Molecular Nutrition Research, Interdisciplinary Research Center, Justus-Liebig-University, Giessen, Germany

    Marco Schumacher & Uwe Wenzel

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  1. Bettina M. Kaminski
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Correspondence to Sandra Ulrich.

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Kaminski, B.M., Weigert, A., Brüne, B. et al. Sulforaphane potentiates oxaliplatin-induced cell growth inhibition in colorectal cancer cells via induction of different modes of cell death. Cancer Chemother Pharmacol 67, 1167–1178 (2011). https://doi.org/10.1007/s00280-010-1413-y

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