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Cancer Chemotherapy and Pharmacology

, Volume 67, Issue 5, pp 1167–1178 | Cite as

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

  • Bettina M. Kaminski
  • Andreas Weigert
  • Bernhard Brüne
  • Marco Schumacher
  • Uwe Wenzel
  • Dieter Steinhilber
  • Jürgen Stein
  • Sandra UlrichEmail author
Original Article

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.

Keywords

Sulforaphane Oxaliplatin Colorectal cancer Cell growth Apoptosis 

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

Notes

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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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Bettina M. Kaminski
    • 1
  • Andreas Weigert
    • 3
  • Bernhard Brüne
    • 3
  • Marco Schumacher
    • 4
  • Uwe Wenzel
    • 4
  • Dieter Steinhilber
    • 1
  • Jürgen Stein
    • 1
    • 2
  • Sandra Ulrich
    • 1
    Email author
  1. 1.Institute of Pharmaceutical ChemistryBiozentrum, Goethe UniversityFrankfurt am MainGermany
  2. 2.Department of Internal MedicineKatharina Kasper HospitalFrankfurt am MainGermany
  3. 3.Institute of Biochemistry I/ZAFESGoethe UniversityFrankfurt am MainGermany
  4. 4.Molecular Nutrition Research, Interdisciplinary Research CenterJustus-Liebig-UniversityGiessenGermany

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