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Differential behaviors of trastuzumab-sensitive and -resistant SKBR3 cells treated with menadione reveal the involvement of Notch1/Akt/FOXO1 signaling elements

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Abstract

Given that HER2 serves as a putative target for therapy in HER2-positive breast cancer, intrinsic and/or acquired resistance to trastuzumab (T) has been proposed to be the major obstacle in treatments. In addition, chemoresistance is commonly attributed to increased antioxidant capacity. In that regard, we evaluated the effect of menadione (M) alone and/or its combination with trastuzumab on proliferation, intracellular GSH and ROS contents as well as HER2 and Notch1 signaling pathways in both trastuzumab-resistant (SKBR3R) and -sensitive SKBR3 (SKBR3S) cells. In spite of increased level of ROS and reduced level of GSH in M-treated SKBR3S cells, M-treated SKBR3R cells showed a decreased content of ROS and GSH compared to untreated cells. However, M/T co-treatment of SKBR3 cells indicated no effect on ROS content, while decreased the level of GSH compared to untreated control cells. Based on the extent of apoptosis, colony formation and wound healing assays, M alone, and/or in combination with T had a stronger inhibitory effect on proliferation of SKBR3R cells relative to SKBR3S cells. These effects might be due to the stronger effects of M and/or M/T on downregulation of p-Akt, Hes1, NICD, and upregulation of FOXO1 among SKBR3R cells relative to the sensitive SKBR3 cells. These findings would certainly shed light on some of the signaling factors involved in induction of trastuzumab resistance and would be of value in designing more efficient chemosensitization strategies.

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Abbreviations

DMSO:

Dimethyl sulfoxide

DCFH:

2′,7′-Dichlorofluorescein diacetate

DTT:

Dithiothreitol

AO/EtBr:

Acridine orange/Ethidium bromide

GSH:

Reduced Glutathione

MCF-7:

Human breast cancer cell line

M:

Menadione

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NAC:

N-Acetylcysteine

NICD:

Notch1 intracellular domain

PMS:

Phenazine methosulphate

ROS:

Reactive oxygen species

R or SKBR3R :

Trastuzumab-resistant SKBR3 cells

S or SKBR3S :

Trastuzumab-sensitive SKBR3 cells

T:

Trastuzumab

T/M:

Trastuzumab/menadione combination

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Acknowledgments

The authors appreciate the financial support of this investigation by the Research Council of University of Tehran.

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Authors and Affiliations

  1. Institute of Biochemistry and Biophysics, University of Tehran, P. O. Box 13145-1384, Tehran, Iran

    Soraya Sajadimajd & Razieh Yazdanparast

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  1. Soraya Sajadimajd
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  2. Razieh Yazdanparast
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Correspondence to Razieh Yazdanparast.

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Sajadimajd, S., Yazdanparast, R. Differential behaviors of trastuzumab-sensitive and -resistant SKBR3 cells treated with menadione reveal the involvement of Notch1/Akt/FOXO1 signaling elements. Mol Cell Biochem 408, 89–102 (2015). https://doi.org/10.1007/s11010-015-2485-0

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  • DOI: https://doi.org/10.1007/s11010-015-2485-0

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