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EBP50 exerts tumor suppressor activity by promoting cell apoptosis and retarding extracellular signal-regulated kinase activity

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Abstract

The expression of Ezrin-radixin-moesin-binding phosphoprotein-50 (EBP50) and the intragenic mutation of the ebp50 gene have been reported to correlate with human breast cancer development, but the exact impacts on breast cancer development and its molecular mechanism are not fully understood. In this study, we investigate the potential function of EBP50 through over-expression in the breast cancer cell line, MDA-MB-231, which has low EBP50 protein expression levels. The effects of EBP50 over-expression on cellular proliferation, anchorage-independent growth and apoptosis were examined. In addition, the activity of extracellular signal-regulated kinase (ERK) was also determined. Our results show that a decrease of cellular proliferation and attenuation of colony-forming ability were evident in MDA-MB-231 cells stably transfected with an EBP50 expressing plasmid (EBP-231) when compared with control cells. There was also a statistically significant increase in spontaneous apoptosis in EBP-231 cells accompanied by an attenuation in ERK activity. Altogether, our results suggest that restoring EBP50 expression could suppress breast cancer cell proliferation by promoting cell apoptosis and inhibiting ERK activity, and that EBP50 may be a target for development of diagnostics and therapeutics in breast cancer.

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Abbreviations

EBP50:

Ezrin-radixin-moesin-binding phosphoprotein-50

NHERF1:

Na+/H+ exchanger regulatory factor 1

ER:

Estrogen receptor

ERK:

The mitogen-activated protein kinases extracellular signal-regulated kinase

PDZ:

Postsynaptic density-95/Discs Large/ZO-1

GPCR:

G protein-coupled receptor

PTEN:

Phosphatase and tensin homolog deleted on chromosome 10

NF2:

Neurofibromatosis 2

SYK:

Spleen tyrosine kinase

PDGFR:

Platelet-derived growth factor receptor

EGFR:

Epidermal growth factor receptor

ATCC:

American type culture collection

DMEM:

Dulbecco’s modified Eagle’s medium

PI:

Propidium iodide

ANOVA:

Analysis of variance

FBS:

Fetal bovine serum

CCK-8:

Cell counting kit-8

HA:

Hemagglutinin

MAPK:

The mitogen-activated kinase

MEF:

Mouse embryonic fibroblast

STAT1:

Signal transducers and activators of transcription 1

PI3K:

PI-3′-OH kinase

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Acknowledgments

This work was supported by the National Natural Science Foundation of the People’s Republic of China (30772573; 30973405; 30900247), the Natural Science Foundation of Beijing (7082009) and the New Century Excellent Talents in University of China (NCET-06-0184), the Beijing Nova program (2008B58), the fund for Excellent Talents in Beijing (20071D0501800253). Screening of cells stably over-expressing EBP50 by Peng Chen and Kun Liu, and the manuscript revision by Prof. Dr. Chuan-Yuan Li are highly appreciated.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Capital Medical University, No. 10 Xitoutiao, You An Men, 100069, Beijing, People’s Republic of China

    Jun-Fang Zheng, Li-Cui Sun, Hua Liu, Yan Huang, Yang Li & Junqi He

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  1. Jun-Fang Zheng
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  2. Li-Cui Sun
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  3. Hua Liu
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  4. Yan Huang
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  5. Yang Li
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  6. Junqi He
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Corresponding author

Correspondence to Junqi He.

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J. Zheng and L. Sun contributed equally to this work.

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Zheng, JF., Sun, LC., Liu, H. et al. EBP50 exerts tumor suppressor activity by promoting cell apoptosis and retarding extracellular signal-regulated kinase activity. Amino Acids 38, 1261–1268 (2010). https://doi.org/10.1007/s00726-009-0437-2

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  • DOI: https://doi.org/10.1007/s00726-009-0437-2

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