Skip to main content

Advertisement

SpringerLink
Log in

A critical role of Sp1 transcription factor in regulating the human Ki-67 gene expression

  • Research Article
  • Published:
Tumor Biology

Abstract

Ki-67 plays a crucial role in cell proliferation as well as maintenance or regulation of cell division. The mechanism governing the Ki-67 gene expression remains unknown. Thus, we cloned the core promoter of the human Ki-67 gene and further investigated its transcriptional regulation. The putative Sp1 binding sites were confirmed by electrophoretic mobility shift assay together with an anti-Sp1 antibody-mediated supershift assay. Deletion mutagenesis and firefly luciferase reporter gene assay demonstrated the essential contribution of Sp1 on transcriptional activation of the Ki-67 gene. In this study, we first confirm that there are three Sp1 binding sites in the Ki-67 core promoter. Two Sp1 sites (one at position −159 to −145 nt and the other at position −14 to +12 nt) are mainly involved in transcriptional regulation of the Ki-67 gene. Overexpression of Sp1 can enhance the Ki-67 promoter activity. However, down-regulation of Sp1 expression using siRNA-Sp1 and mithramycin effectively inhibits the Ki-67 gene transcription. Our results suggest that Sp1 is essential for basal promoter activity of the human Ki-67 gene. Inhibition of the Ki-67 transcriptional activity through abolishment of Sp1 may provide the useful prospect for gene therapy.

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
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

Sp1:

Specificity protein 1

RT-PCR:

Reverse transcription polymerase chain reaction

siRNA:

Small interfering RNA

EMSA:

Electrophoretic mobility shift assays

SV40:

Simian virus 40 promoter

HRP:

Horseradish peroxidase

TBST:

Tris-buffered saline Tween-20

References

  1. Duchrow M, Schlueter C, Wohlenberg C, Flad HD, Gerdes J. Molecular characterization of the gene locus of the human cell proliferation-associated nuclear protein defined by monoclonal antibody Ki-67. Cell Prolif. 1996;29:1–12.

    Article  CAS  PubMed  Google Scholar 

  2. MacCallum DE, Hall PA. Biochemical characterization of pKi67 with the identification of a mitotic-specific form associated with hyperphosphorylation and altered DNA binding. Exp Cell Res. 1999;252:186–98.

    Article  CAS  PubMed  Google Scholar 

  3. Schlüter C, Duchrow M, Wohlenberg C, Becker MH, Key G, Flad HD, et al. The cell proliferation associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins. J Cell Biol. 1993;123:513–22.

    Article  PubMed  Google Scholar 

  4. MaCallum DE, Hall PA. Biochemical characterization of pKi-67 with the identification of a mitotic-specific form association with hyperphosphorylation and altered DNA binding. Exp Cell Res. 1991;52:186–98.

    Google Scholar 

  5. Kausch I, Jiang H, Ewerdwalbesloh N, Doehn C, Kruger S, Sczakiel G, et al. Inhibition of Ki-67 in a renal cell carcinoma severe combined immunodeficiency disease mouse model is associated with induction of apoptosis and tumour growth inhibition. BJU Int. 2005;95:416–20.

    Article  CAS  PubMed  Google Scholar 

  6. Dynan WS, Tjian R. Isolation of transcription factors that discriminate between different promoters recongnized by RNA polymerase II. Cell. 1983;32:669–80.

    Article  CAS  PubMed  Google Scholar 

  7. Dynan WS, Tjian R. The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter. Cell. 1983;35:79–87.

    Article  CAS  PubMed  Google Scholar 

  8. Suske G. The Sp-family of transcription factors. Gene. 1999;238:291–300.

    Article  CAS  PubMed  Google Scholar 

  9. Kadonaga JT, Carner KR, Masiarz FR, Tjian R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell. 1987;51:1079–90.

    Article  CAS  PubMed  Google Scholar 

  10. Endl E, Gerdes J. Post-translational modifications of the Ki-67 protein coincide with two major check points during mitosis. J Cell Physiol. 2000;182:371–80.

    Article  CAS  PubMed  Google Scholar 

  11. Scholzen T, Gerdes J. The Ki-67 protein: from the known and the unknown. J Cell Physiol. 2000;182:311–22.

    Article  CAS  PubMed  Google Scholar 

  12. Takagi M, Matsuoka Y, Kurihara T, Yoneda Y. Chmadrin: a novel Ki-67 antigen-related perichromosomal protein possibly implicated in higher order chromatin structure. J Cell Sci. 1999;112:2463–72.

    CAS  PubMed  Google Scholar 

  13. Yang X, Su K, Roos MD, Chang Q, Paterson AJ, Kudlow JE. O-linkage of N-acetylglucosamine to Sp1 activation domain inhibits its transcriptional capability. Proc Natl Acad Sci. 2001;98:6611–6.

    Article  CAS  PubMed  Google Scholar 

  14. Su K, Roos MD, Yang X, Han I, Paterson AJ, Kudlow JE. An N-terminal region of Sp1 targets its proteasome dependent degradation in vitro. J Biol Chem. 1999;274:15194–202.

    Article  CAS  PubMed  Google Scholar 

  15. Safe S, Abdelrahim M. Sp transcription factor family and its role in cancer. Eur J Cancer. 2005;41:2438–48.

    Article  CAS  PubMed  Google Scholar 

  16. Safe S, Kim K. Nuclear receptor-mediated transactivation through interaction with Sp proteins. Prog Nucleic Acid Res Mol Biol. 2004;77:1–36.

    Article  CAS  PubMed  Google Scholar 

  17. Bouwman P, Philipsen S. Regulation of the activity of Sp1-related transcription factors. Mol Cell Endocrinol. 2002;195:27–38.

    Article  CAS  PubMed  Google Scholar 

  18. Li L, He S, Sun JM, Davie JR. Gene regulation by Sp1 and Sp3. Biochem Cell Biol. 2004;82:460–71.

    Article  CAS  PubMed  Google Scholar 

  19. Barth N, Langmann T, Scholmerich J, Schmitz G, Schaffler A. Identification of regulatory elements in the human adipose most abundant gene transcript-1 (apM-1) promoter: role of SP1/SP3 and TNF-alpha as regulatory pathways. Diabetologia. 2002;45:1425–33.

    CAS  PubMed  Google Scholar 

  20. Wang LW, Wei DY, Huang SY, Peng ZH, Le XD, Wu TT, et al. Transcription factor Sp1 expression is a significant predictor of survival in human gastric cancer. Clin Cancer Res. 2003;9:6371–80.

    CAS  PubMed  Google Scholar 

  21. Shi Q, Le XD, Abbruzzese JL, Peng ZH, Qian CN, Tang HM, et al. Constitutive Sp1 activity is essential for differential constitutive expression of vascular endothelial growth factor in human pancreatic adenocarcinoma. Cancer Res. 2001;61:4143–54.

    CAS  PubMed  Google Scholar 

  22. Chiefari E, Brunetti A, Arturi F, Bidart JM, Russo D, Schlumberger M, et al. Increased expression of AP2 and Sp1 transcription factors in human thyroid tumours: a role in NIS expression regulation. BMC Cancer. 2002;2:35.

    Article  PubMed  Google Scholar 

  23. Hosoi Y, Watanabe T, Nakagawa K, Matsumoto Y, Enomoto A, Morita A, et al. Up-regulation of DNA-dependent protein kinase activity and Sp1 in colorectal cancer. Int J Oncol. 2004;25:461–8.

    CAS  PubMed  Google Scholar 

  24. Jang SI, Steinert PM. Loricrin expression in cultured human keratinocytes is controlled by a complex interplay between transcription factors of the Sp1, CREB, AP1, and AP2 families. J Biol Chem. 2002;277:42268–79.

    Article  CAS  PubMed  Google Scholar 

  25. Suzuki T, Kimura A, Nagai R, Horikoshi M. Regulation of interaction of the acetyltransferase region of p300 and the DNA-binding domain of Sp1 on and through DNA binding. Genes Cells. 2000;5:29–41.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This project is supported by grants from the National Natural Science Foundation of China (nos. 30873021 and 30972976) and the Science and Technology Department of Jiangsu Province (no. BK 2010177).

Conflicts of interest

None declared.

Author information

Authors and Affiliations

  1. Laboratory of Biological Cancer Therapy, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, Jiangsu, 221002, People’s Republic of China

    Hui Tian, Wang Li, Fei-Fei Chen, Jie-Hui Di, Bao-Fu Zhang, Dong-Sheng Pei & Jun-Nian Zheng

  2. Department of Clinical Laboratory, Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221001, People’s Republic of China

    Guo-Wei Qian & Ping Ma

Authors
  1. Hui Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guo-Wei Qian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Fei-Fei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jie-Hui Di
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bao-Fu Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dong-Sheng Pei
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ping Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jun-Nian Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun-Nian Zheng.

Additional information

Hui Tian and Guo-Wei Qian contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tian, H., Qian, GW., Li, W. et al. A critical role of Sp1 transcription factor in regulating the human Ki-67 gene expression. Tumor Biol. 32, 273–283 (2011). https://doi.org/10.1007/s13277-010-0119-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-010-0119-4

Keywords

Search

Navigation