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AP4 activates cell migration and EMT mediated by p53 in MDA-MB-231 breast carcinoma cells

Molecular and Cellular Biochemistry volume 407, pages 57–68 (2015)Cite this article

Abstract

Tumor metastasis is the primary cause of mortality in most cancer patients. Before disassociation from the tumors, most of malignant tumor cells undergo the epithelial–mesenchymal transition to break away from the adhesions between the cells and the surrounding extracellular matrix. Recently, activating enhancer-binding protein (AP4) has been shown to be a mediator of EMT in colorectal cancer and high level of AP4 correlates with poor prognosis in cancer patients. It has been found that AP4 upregulates the genes involved in EMT and cell proliferation in colorectal cancer cells and that the aggressive human breast cancer cells MDA-MB-231 are highly metastatic. Therefore, we tested the hypothesis that AP4 may also affect cell migration and EMT in this cell type. Three different assays, including the wound-healing assay, the Boyden chamber assay, and the cell tracking assay, were employed to confirm that AP4 activated both cell migration and invasion. Immunofluorescence staining and Western blot analysis revealed that the cells underwent EMT when AP4 was upregulated. In contrast, overexpression of dominant-negative AP4, lacking the DNA-binding domain, inactivated the DNA-binding ability of endogenous AP4 and led to lower cell motility. Furthermore, we found that AP4 enhanced p53 expression at both transcriptional and translational levels. Knockdown of p53 by siRNA significantly diminished the activation of cell migration by AP4, indicating that AP4 can regulate cell migration via the activity of p53.

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Abbreviations

AP4:

Activating enhancer-binding-4

DN-AP4:

Dominant negative AP4

bHLH:

Basic helix-loop-helix

SV40:

Simian virus 40

MMP-9:

Matrix metalloproteinase-9

IGFBP-2:

Insulin-like growth factor-binding protein 2

mdm2 :

Murine double minute clone 2

EMT:

Epithelial–mesenchymal transition

MET:

Mesenchymal–epithelial transition

DOX:

Doxycycline

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Acknowledgments

This work was supported by the Seed Grant of City University of Hong Kong (Project no. 7004028).

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  1. Department of Biomedical Sciences, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong, Hong Kong

    Shaopeng Chen & Sung-Kay Chiu

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  1. Shaopeng Chen
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Correspondence to Sung-Kay Chiu.

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Chen, S., Chiu, SK. AP4 activates cell migration and EMT mediated by p53 in MDA-MB-231 breast carcinoma cells. Mol Cell Biochem 407, 57–68 (2015). https://doi.org/10.1007/s11010-015-2454-7

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Keywords

  • AP4
  • Cell migration
  • Mutant p53
  • EMT
  • Cell invasion
  • Metastatic MDA-MB-231 cells