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Notch2 activation by benzyl isothiocyanate impedes its inhibitory effect on breast cancer cell migration

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Abstract

Benzyl isothiocyanate (BITC) is a promising anticancer constituent of edible cruciferous vegetables with in vivo efficacy against chemically induced as well as oncogene-driven breast cancer in experimental rodents. However, the mechanism underlying anticancer effect of BITC is not fully understood. This study was undertaken to determine the role of Notch signaling in anticancer responses to BITC as this pathway is often hyperactive in human breast cancer. Exposure of MCF-7, MDA-MB-231, and SUM159 human breast cancer cells to pharmacologic concentrations of BITC (2.5 and 5 μM) resulted in cleavage (activation) of Notch1, Notch2, and Notch4, which was accompanied by induction of γ-secretase complex components Presenilin1 and/or Nicastrin. The BITC-mediated cleavage of Notch was associated with its transcriptional activation as revealed by RBP-Jk and Hes-1A/B luciferase reporter assays. Inhibition of cell migration or cell viability resulting from BITC exposure was not influenced by pharmacological suppression of Notch1 using a γ-secretase inhibitor or RNA interference of Notch1 as well as Notch4. On the other hand, the BITC-mediated inhibition of cell migration, but not cell viability, was significantly augmented by siRNA and shRNA knockdown of Notch2 protein. Furthermore, the BITC-mediated inhibition of MDA-MB-231 xenograft growth in vivo was associated with a significant increase in nuclear levels of cleaved Notch2 and Hes-1 proteins. In conclusion, the results of this study indicate that (a) BITC treatment activates Notch2 in cultured and xenografted human breast cancer cells, and (b) Notch2 activation impedes inhibitory effect of BITC on cell migration.

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Abbreviations

BITC:

Benzyl isothiocyanate

EMT:

Epithelial-mesenchymal transition

DAPT:

N-[N-(3,5-difluorophenacetyl-L-alanyl)]-S-phenylglycine-t-butyl ester

siRNA:

Small interfering RNA

shRNA:

Small hairpin RNA

DMSO:

Dimethyl sulfoxide

uPA:

Urokinase-type plasminogen activator

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Conflict of interest

None of the authors has any conflict of interest. This study was supported by the US PHS grant RO1 CA129347-05 awarded by the National Cancer Institute.

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Authors and Affiliations

  1. Department of Pharmacology & Chemical Biology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, 2.32A Hillman Cancer Center Research Pavilion, 5117 Centre Avenue, Pittsburgh, PA, 15213, USA

    Su-Hyeong Kim, Anuradha Sehrawat & Shivendra V. Singh

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  1. Su-Hyeong Kim
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  2. Anuradha Sehrawat
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  3. Shivendra V. Singh
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Correspondence to Shivendra V. Singh.

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Kim, SH., Sehrawat, A. & Singh, S.V. Notch2 activation by benzyl isothiocyanate impedes its inhibitory effect on breast cancer cell migration. Breast Cancer Res Treat 134, 1067–1079 (2012). https://doi.org/10.1007/s10549-012-2043-3

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