Skip to main content
SpringerLink
Log in

Transcriptional regulation of human MUC4 gene: identification of a novel inhibitory element and its nuclear binding protein

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

The human mucin 4 (MUC4) is aberrantly expressed in pancreatic adenocarcinoma and tumor cell lines, while remaining undetectable in normal pancreas, indicating its important role in pancreatic cancer development. Although its transcriptional regulation has been investigated in considerable detail, some important elements remain unknown. The aim of the present study was to demonstrate the existence of a novel inhibitory element in the MUC4 promoter and characterize some of its binding proteins. By luciferase reporter assay, we located the inhibitory element between nucleotides −2530 and −2521 in the MUC4 promoter using a series of deletion and mutant reporter constructs. Electrophoretic mobility shift assay (EMSA) with Bxpc-3 cell nuclear extracts revealed that one protein or protein complex bind to this element. The proteins binding to this element were purified and identified as Yin Yang 1 (YY1) by mass spectrometry. Supershift assay and chromatin immunoprecipitation (ChIP) assay confirmed that YY1 binds to this element in vitro and in vivo. Moreover, transient YY1 overexpression significantly inhibited MUC4 promoter activity and endogenous MUC4 protein expression. In conclusion, we reported here a novel inhibitory element in the human MUC4 promoter. This provides additional data on MUC4 gene regulation and indicates that YY1 may be a potential target for abnormal MUC4 expression.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Porchet N, Nguyen VC, Dufosse J et al (1991) Molecular cloning and chromosomal localization of a novel human tracheo-bronchial mucin cDNA containing tandemly repeated sequences of 48 base pairs. Biochem Biophys Res Commun 175:414–422

    Article  PubMed  CAS  Google Scholar 

  2. Andrianifahanana M, Moniaux N, Schmied BM et al (2001) Mucin (MUC) gene expression in human pancreatic adenocarcinoma and chronic pancreatitis: a potential role of MUC4 as a tumor marker of diagnostic significance. Clin Cancer Res 7:4033–4040

    PubMed  CAS  Google Scholar 

  3. Iacobuzio-Donahue CA, Ashfaq R, Maitra A et al (2003) Highly expressed genes in pancreatic ductal adenocarcinomas: a comprehensive characterization and comparison of the transcription profiles obtained from three major technologies. Cancer Res 63:8614–8622

    PubMed  CAS  Google Scholar 

  4. Swartz MJ, Batra SK, Varshney GC et al (2002) MUC4 expression increases progressively in pancreatic intraepithelial neoplasia. Am J Clin Pathol 117:791–796

    Article  PubMed  Google Scholar 

  5. Zhu Y, Zhang JJ, Zhu R et al (2011) The increase in the expression and hypomethylation of MUC4 gene with the progression of pancreatic ductal adenocarcinoma. Med Oncol 28:S175–S184

    Article  PubMed  Google Scholar 

  6. Singh AP, Moniaux N, Chauhan SC et al (2004) Inhibition of MUC4 expression suppresses pancreatic tumor cell growth and metastasis. Cancer Res 64:622–630

    Article  PubMed  CAS  Google Scholar 

  7. Perrais M, Pigny P, Ducourouble MP et al (2001) Characterization of human mucin gene MUC4 promoter: importance of growth factors and proinflammatory cytokines for its regulation in pancreatic cancer cells. J Biol Chem 276:30923–30933

    Article  PubMed  CAS  Google Scholar 

  8. Andrianifahanana M, Singh AP, Nemos C et al (2007) IFN-γ-induced expression of MUC4 in pancreatic cancer cells is mediated by STAT-1 upregulation: a novel mechanism for IFN-gamma response. Oncogene 26:7251–7261

    Article  PubMed  CAS  Google Scholar 

  9. Mejías-Luque R, Peiró S, Vincent A et al (2008) IL-6 induces MUC4 expression through gp130/STAT3 pathway in gastric cancer cell lines. Biochim Biophys Acta 1783:1728–1736

    Article  PubMed  Google Scholar 

  10. Bai CH, Song SY, Kim YD (2007) Effect of glucocorticoid on the MUC4 gene in nasal polyps. Laryngoscope 117:2169–2173

    Article  PubMed  CAS  Google Scholar 

  11. Vincent A, Ducourouble MP, Van Seuningen I (2008) Epigenetic regulation of the human mucin gene MUC4 in epithelial cancer cell lines involves both DNA methylation and histone modifications mediated by DNA methyltransferases and histone deacetylases. FASEB J 22:3035–3045

    Article  PubMed  CAS  Google Scholar 

  12. Yamada N, Nishida Y, Tsutsumida H et al (2009) Promoter CpG methylation in cancer cells contributes to the regulation of MUC4. Br J Cancer 100:344–351

    Article  PubMed  CAS  Google Scholar 

  13. Zhang JJ, Zhang XF, Zhu Y et al (2010) Transcriptional regulation of human mucin gene MUC4 in pancreatic cancer cells. Mol Biol Rep 37:2797–2802

    Article  PubMed  CAS  Google Scholar 

  14. Fauquette V, Perrais M, Cerulis S et al (2005) The antagonistic regulation of human MUC4 and ErbB-2 genes by the Ets protein PEA3 in pancreatic cancer cells: implications for the proliferation/differentiation balance in the cells. Biochem J 386:35–45

    Article  PubMed  CAS  Google Scholar 

  15. Piessen G, Jonckheere N, Vincent A et al (2007) Regulation of the human mucin MUC4 by taurodeoxycholic and taurochenodeoxycholic bile acids in oesophageal cancer cells is mediated by hepatocyte nuclear factor 1α. Biochem J 402:81–91

    Article  PubMed  CAS  Google Scholar 

  16. Jonckheere N, Vincen A, Perrais M et al (2007) The human mucin MUC4 is transcriptionally regulated by caudal-related homeobox, hepatocyte nuclear factors, forkhead box A, and GATA endodermal transcription factors in epithelial cancer cells. J Biol Chem 282:22638–22650

    Article  PubMed  CAS  Google Scholar 

  17. Fauquette V, Aubert S, Groux-Degroote S et al (2007) Transcription factor AP-2α represses both the mucin MUC4 expression and pancreatic cancer cell proliferation. Carcinogenesis 28:2305–2312

    Article  PubMed  CAS  Google Scholar 

  18. Shi Y, Seto E, Chang LS et al (1991) Transcriptional repression by YY1, a human GLI-Kruppel-related protein, and relief of repression by adenovirus E1A protein. Cell 67:377–388

    Article  PubMed  CAS  Google Scholar 

  19. Thomas MJ, Seto E (1999) Unlocking the mechanisms of transcription factor YY1: are chromatin modifying enzymes the key? Gene 236:197–208

    Article  PubMed  CAS  Google Scholar 

  20. Shi Y, Lee JS, Galvin KM (1997) Everything you have ever wanted to know about Yin Yang 1. Biochim Biophys Acta 1332:F49–F66

    PubMed  CAS  Google Scholar 

  21. Srinivasan L, Atchison ML (2004) YY1 DNA binding and PcG recruitment requires CtBP. Genes Dev 18:2596–2601

    Article  PubMed  CAS  Google Scholar 

  22. Sui G, el Affar B, Shi Y et al (2004) Yin Yang 1 is a negative regulator of p53. Cell 117:859–872

    Article  PubMed  CAS  Google Scholar 

  23. Yang WM, Inouye C, Zeng Y et al (1996) Transcriptional repression by YY1 is mediated by interaction with a mammalian homolog of the yeast global regulator RPD3. Proc Natl Acad Sci USA 93:12845–12850

    Article  PubMed  CAS  Google Scholar 

  24. Schug J, Schuller WP, Kappen C et al (2005) Promoter features related to tissue specificity as measured by Shannon entropy. Genome Biol 6:R33

    Article  PubMed  Google Scholar 

  25. Seto E, Lewis B, Shenk T (1993) Interaction between transcription factors Sp1 and YY1. Nature 365:462–464

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported in part by the Grants from the National Natural Science Foundation of China (81001079, 30972912 and 81101802), the Program for Development of Innovative Research Team in the First Affiliated Hospital of NJMU, the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), the Natural Science Foundation of Zhejiang Province (Y2110310), and the Science and Technology Project of Wenzhou City (Y20100243).

Author information

Authors and Affiliations

  1. Department of General Surgery, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, People’s Republic of China

    Jing-Jing Zhang, Yi Zhu, Ze-Kuan Xu & Yi Miao

  2. Jiangsu Province Academy of Clinical Medicine, Institute of Tumor Biology, 300 Guangzhou Road, Nanjing, 210029, People’s Republic of China

    Jing-Jing Zhang, Yi Zhu, Ze-Kuan Xu & Yi Miao

  3. Department of Biochemistry, Wenzhou Medical College, Wenzhou, 325035, People’s Republic of China

    Xiong-Fei Zhang

  4. Transfusion Laboratory, Jiangsu Province Blood Center, Nanjing, 210029, People’s Republic of China

    Wen-Biao Liang

  5. The First School of Clinical Medicine, Nanjing Medical University, 140 Hanzhong Road, Nanjing, 210029, People’s Republic of China

    Kun-Ling Xie, Jin-Qiu Tao & Yun-Peng Peng

Authors
  1. Jing-Jing Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yi Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiong-Fei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wen-Biao Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kun-Ling Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jin-Qiu Tao
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yun-Peng Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ze-Kuan Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yi Miao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Ze-Kuan Xu or Yi Miao.

Additional information

Jing-Jing Zhang and Yi Zhu contributed equally to this study.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, JJ., Zhu, Y., Zhang, XF. et al. Transcriptional regulation of human MUC4 gene: identification of a novel inhibitory element and its nuclear binding protein. Mol Biol Rep 40, 4913–4920 (2013). https://doi.org/10.1007/s11033-013-2591-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-013-2591-6

Keywords

Search

Navigation