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JMJD2A-dependent silencing of Sp1 in advanced breast cancer promotes metastasis by downregulation of DIRAS3

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An Erratum to this article was published on 23 February 2016

Abstract

Specificity protein 1(Sp1) is a ubiquitous transcription factor and is highly expressed in breast cancer. However, its expression pattern and role in breast cancer progression remain unclear. The purpose of this study is to examine the expression pattern of Sp1 and determine its role in breast cancer progression. Immunohistochemistry (IHC) was performed on breast cancer tissues to reveal the expression pattern of Sp1. Spearman rank correlation was used for clinical statistics. Gene and protein expressions were monitored by IHC analysis, quantitative polymerase chain reaction, and Western blot. Wound-healing and Transwell assays were conducted to assess the role of Sp1 in breast cancer. Co-immunoprecipitation, deletion mutagenesis, chromatin immunoprecipitation, and dual luciferase reporter gene assays were used for investigation of the regulatory network. Sp1 expression was downregulated in late stage breast cancer and in highly invasive breast cancer cell lines. Expression of Sp1 was negatively correlated with TNM staging (P = 0.002) and metastasis status (P = 0.023). Overexpression of Sp1 inhibited breast cancer cell migratory and invasive abilities, whereas knockdown of GTP-binding RAS-like 3 (DIRAS3, also known as ARHI, NOEY2) attenuated the inhibitory effects. Moreover, re-expression of DIRAS3 abolished Sp1 knockdown-mediated cell migration and invasion. Jumonji domain containing 2A (JMJD2A) inhibited Sp1 autoregulation and explains Sp1 expression pattern in breast cancer. Sp1 negatively regulated breast cancer metastasis by transcriptional activation of DIRAS3. Inhibition of Sp1 autoregulation by JMJD2A contributed to Sp1 expression pattern in breast cancer. Our findings provided evidence that targeted therapy against Sp1 might be useful in early stage breast cancer. However, in late stages, development of Sp1 activator may be more promising for breast cancer treatments.

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Abbreviations

Sp1:

Specificity protein 1

DIRAS3:

GTP-binding RAS-like 3

JMJD2A:

Jumonji containing domain 2A

qRT-PCR:

Quantitative real-time polymerase chain reaction

ChIP:

Chromatin immunoprecipitation

Co-IP:

Co-immunoprecipitation

ATCC:

American type culture collection

DMEM:

Dulbecco’s modified eagle medium

FBS:

Fetal bovine serum

IHC:

Immunohistochemistry

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81172897). We thank Dr. Ralf Janknecht from Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine and Dr. Hsing-Jien Kung from UC Davis Cancer Center for JMJD2A expression plasmids, and Dr. Jun Fujita from Department of Clinical Molecular Biology, Kyoto University for the Sp1 plasmid. We also feel great thanks for Dr. Zhao-Hui Wu from the Health Science Center, University of Tennessee and Dr. Joseph Washington from Laboratory for Implantable Materials and Biomaterials, Department of Mechanical Engineering, University of Maryland, Baltimore County for their helps in polishing this article and Dr. Zude Xu from Huashan Hospital, Shanghai, China for the clinical specimen and his professional help in pathological diagnosis. Dr. Liliang Li acknowledges the support from the State Scholarship Fund of China Scholarship Council for his visit to the University of Maryland School of Medicine through the joint Ph.D student programme (student No.: 201306100058), where part of this work was conducted.

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Xuhui District, Shanghai, 200032, People’s Republic of China

    Liliang Li, Yuhua Li, Yiwen Shen, Jianhui Xie, Aimin Xue, Ziqin Zhao, Mingchang Zhang, Beixu Li & Jieqing Jiang

  2. Department of Pathology, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Xuhui District, Shanghai, 200032, People’s Republic of China

    Pan Gao & Zude Xu

  3. Division of Forensic Pathology, University of Maryland School of Medicine, 900 West Baltimore Street, Baltimore, MD, 21223, USA

    Liliang Li

  4. Forensic Science Center, East China University of Political Science and Law, 112 Huayang Road, Changning District, Shanghai, 200042, People’s Republic of China

    Daming Sun

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  1. Liliang Li
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Corresponding authors

Correspondence to Aimin Xue or Ziqin Zhao.

Additional information

Liliang Li and Pan Gao contributed equally to this work.

Electronic supplementary material

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10549_2014_3083_MOESM1_ESM.tif

Supplementary material 1 (TIFF 2129 kb) Supplementary Fig. 1 mRNA level of Sp1 was negatively associated with that of DIRAS3 and positively associated with that of JMJD2A in MDA-MB-231 cells. a mRNA level of DIRAS3 was elevated in response to increased Sp1 mRNA level. b mRNA level of DIRAS3 was accordingly reduced along with decreased Sp1 mRNA level. c When JMJD2A was upregulated, mRNA level of Sp1 was inversely downregulated. d Knockdown of JMJD2A by specific siRNAs increased mRNA level of Sp1. *P < 0.05. siNC, negative control siRNA

10549_2014_3083_MOESM2_ESM.tif

Supplementary material 2 (TIFF 3174 kb) Supplementary Fig. 2 knockdown of DIRAS3 rescued Sp1 overexpression-suppressed breast cancer migration and invasion. a Sp1 overexpression suppressed MDA-MB-231 cells wound recovery process, whereas knockdown of DIRAS3 in Sp1-overexpressed cells alleviated the suppression mediated by Sp1 overexpression. b transwell assays confirmed that knockdown of DIRAS3 deprived the suppression of breast cancer migration and invasion by Sp1 overexpression

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Li, L., Gao, P., Li, Y. et al. JMJD2A-dependent silencing of Sp1 in advanced breast cancer promotes metastasis by downregulation of DIRAS3. Breast Cancer Res Treat 147, 487–500 (2014). https://doi.org/10.1007/s10549-014-3083-7

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  • DOI: https://doi.org/10.1007/s10549-014-3083-7

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