Skip to main content

Advertisement

SpringerLink
Log in

Promoter-associated endogenous and exogenous small RNAs suppress human bladder cancer cell metastasis by activating p21 CIP1/WAF1 expression

  • Original Article
  • Published:
Tumor Biology

Abstract

Accumulating data suggest that micro RNAs (miRNAs) or double-stranded RNAs (dsRNAs) can activate gene expression by targeting promoters. The cyclin-dependent kinase inhibitor p21 CIP1/WAF1 (p21) has also been shown to suppress epithelial-mesenchymal transition (EMT) which plays a crucial role in the early stage of tumor metastases and invasiveness. In a previous study, we have reported that miR-370-5p is low-expressed in bladder cancer (BCa) tissues and cell lines. Here, we identified that miR-370-5p and sequence homology dsRNA (dsP21-555) fully complementary to promoter hold the potent abilities to induce p21 expression. Moreover, transfection of miR-370-5p or dsP21-555 into BCa cells remarkably inverts EMT-associated genes (increases epithelial cell makers E-cadherin and β-catenin, and decreases mesenchymal cell markers ZEB1 and Vimentin) expression mainly via regulating p21 expression. Besides, through manipulating p21, both the candidates can retard BCa cell migration and invasion. In summary, our results provide evidence that both endogenous and exogenous small RNAs may function to induce p21 expression by interacting with the similar promoter region and impede BCa metastasis.

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
Fig. 7

Similar content being viewed by others

References

  1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA: Cancer J Clin. 2011;61:69–90.

    Google Scholar 

  2. Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer J Int Du cancer. 2010;127:2893–917.

    Article  CAS  Google Scholar 

  3. Burger M, Catto JW, Dalbagni G, Grossman HB, Herr H, Karakiewicz P, et al. Epidemiology and risk factors of urothelial bladder cancer. Eur Urol. 2013;63:234–41.

    Article  PubMed  Google Scholar 

  4. Kaufman DS, Shipley WU, Feldman AS. Bladder cancer. Lancet. 2009;374:239–49.

    Article  CAS  PubMed  Google Scholar 

  5. Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA: Cancer J Clin. 2014;64:9–29.

    Google Scholar 

  6. Warfel NA, El-Deiry WS. p21waf1 and tumourigenesis: 20 years after. Curr Opin Oncol. 2013;25:52–8.

    Article  CAS  PubMed  Google Scholar 

  7. McConkey DJ, Choi W, Marquis L, Martin F, Williams MB, Shah J, et al. Role of epithelial-to-mesenchymal transition (EMT) in drug sensitivity and metastasis in bladder cancer. Cancer Metastasis Rev. 2009;28:335–44.

    Article  CAS  PubMed  Google Scholar 

  8. Gavert N, Ben-Ze'ev A. Epithelial-mesenchymal transition and the invasive potential of tumors. Trends Mol Med. 2008;14:199–209.

    Article  CAS  PubMed  Google Scholar 

  9. Katsuno Y, Lamouille S, Derynck R. Tgf-β signaling and epithelial-mesenchymal transition in cancer progression. Curr Opin Oncol. 2013;25:76–84.

    Article  CAS  PubMed  Google Scholar 

  10. Li LC, Okino ST, Zhao H, Pookot D, Place RF, Urakami S, et al. Small dsRNAs induce transcriptional activation in human cells. Proc Natl Acad Sci U S A. 2006;103:17337–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Place RF, Li LC, Pookot D, Noonan EJ, Dahiya R. MicroRNA-373 induces expression of genes with complementary promoter sequences. Proc Natl Acad Sci U S A. 2008;105:1608–13.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Kang MR, Yang G, Place RF, Charisse K, Epstein-Barash H, Manoharan M, et al. Intravesical delivery of small activating RNA formulated into lipid nanoparticles inhibits orthotopic bladder tumor growth. Cancer Res. 2012;72:5069–79.

    Article  CAS  PubMed  Google Scholar 

  13. Kosaka M, Kang MR, Yang G, Li LC. Targeted p21WAF1/CIP1 activation by RNAa inhibits hepatocellular carcinoma cells. Nucleic Acid Ther. 2012;22:335–43.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Wei J, Zhao J, Long M, Han Y, Wang X, Lin F, et al. p21WAF1/CIP1 gene transcriptional activation exerts cell growth inhibition and enhances chemosensitivity to cisplatin in lung carcinoma cell. BMC Cancer. 2010;10:632.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Junxia W, Ping G, Yuan H, Lijun Z, Jihong R, Fang L, et al. Double strand RNA-guided endogeneous E-cadherin up-regulation induces the apoptosis and inhibits proliferation of breast carcinoma cells in vitro and in vivo. Cancer Sci. 2010;101:1790–6.

    Article  PubMed  Google Scholar 

  16. Wang C, Chen Z, Ge Q, Hu J, Li F, Hu J, et al. Upregulation of p21 by miRNAs and its implications in bladder cancer cells. FEBS Lett. 2014;588:4654–64.

    Article  CAS  PubMed  Google Scholar 

  17. Miyazaki M, Sakaguchi M, Akiyama I, Sakaguchi Y, Nagamori S, Huh NH. Involvement of interferon regulatory factor 1 and S100C/A11 in growth inhibition by transforming growth factor β1 in human hepatocellular carcinoma cells. Cancer Res. 2004;64:4155–61.

    Article  CAS  PubMed  Google Scholar 

  18. Huang V, Qin Y, Wang J, Wang XL, Place RF, Lin GT, et al. RNAa is conserved in mammalian cells. PLoS One. 2010;5.

  19. Li Q, Li J, Wen T, Zeng W, Peng C, Yan S, et al. Overexpression of HMGB1 in melanoma predicts patient survival and suppression of HMGB1 induces cell cycle arrest and senescence in association with p21 (WAF1/CIP1) up-regulation via a p53-independent, Sp1-dependent pathway. Oncotarget. 2014;5:6387–403.

    Article  PubMed  PubMed Central  Google Scholar 

  20. McConkey DJ, Lee S, Choi W, Tran M, Majewski T, Lee S, et al. Molecular genetics of bladder cancer: emerging mechanisms of tumor initiation and progression. Urol Oncol. 2010;28:429–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Liu M, Casimiro MC, Wang C, Shirley LA, Jiao X, Katiyar S, et al. P21CIP1 attenuates Ras- and c-myc-dependent breast tumor epithelial mesenchymal transition and cancer stem cell-like gene expression in vivo. Proc Natl Acad Sci U S A. 2009;106:19035–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Li XL, Hara T, Choi YG, Subramanian M, Francis P, Bilke S, et al. A p21-ZEB1 complex inhibits epithelial-mesenchymal transition through the microRNA 183-96-182 cluster. Mol Cell Biol. 2014;34:533–50.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Behnsawy HM, Miyake H, Abdalla MA, Sayed MA, Ahmed AEFI, Fujisawa M. Expression of cell cycle-associated proteins in non-muscle-invasive bladder cancer: correlation with Intravesical recurrence following transurethral resection. Urol Oncol-Semin Ori. 2011;29:495–501.

    Article  CAS  Google Scholar 

  24. Tang K, Wang C, Chen Z, Xu H, Ye Z. Clinicopathologic and prognostic significance of p21 (Cip1/Waf1) expression in bladder cancer. Int J Clin Exp Pathol. 2015;8:4999–5007.

    PubMed  PubMed Central  Google Scholar 

  25. Tiguert R, Lessard A, So A, Fradet Y. Prognostic markers in muscle invasive bladder cancer. World J Urol. 2002;20:190–5.

    CAS  PubMed  Google Scholar 

  26. Stein JP, Ginsberg DA, Grossfeld GD, Chatterjee SJ, Esrig D, Dickinson MG, et al. Effect of p21WAF1/CIP1 expression on tumor progression in bladder cancer. J Natl Cancer Inst. 1998;90:1072–9.

    Article  CAS  PubMed  Google Scholar 

  27. Place RF, Wang J, Noonan EJ, Meyers R, Manoharan M, Charisse K, et al. Formulation of small activating RNA into lipidoid nanoparticles inhibits xenograft prostate tumor growth by inducing p21 expression. Mol Ther-Nucl Acids. 2012;1.

  28. Portnoy V, Huang V, Place RF, Li LC. Small RNA and transcriptional upregulation. Wires Rna. 2011;2:748–60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Janowski BA, Younger ST, Hardy DB, Ram R, Huffman KE, Corey DR. Activating gene expression in mammalian cells with promoter-targeted duplex RNAs. Nat Chem Biol. 2007;3:166–73.

    Article  CAS  PubMed  Google Scholar 

  30. Place RF, Noonan EJ, Foldes-Papp Z, Li LC. Defining features and exploring chemical modifications to manipulate RNAa activity. Curr Pharm Biotechnol. 2010;11:518–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Zheng L, Wang L, Gan J, Zhang H. RNA activation: promise as a new weapon against cancer. Cancer Lett. 2014;355:18–24.

    Article  CAS  PubMed  Google Scholar 

  32. Turunen MP, Lehtola T, Heinonen SE, Assefa GS, Korpisalo P, Girnary R, et al. Efficient regulation of VEGF expression by promoter-targeted lentiviral shRNAs based on epigenetic mechanism: a novel example of epigenetherapy. Circ Res. 2009;105:604–9.

    Article  CAS  PubMed  Google Scholar 

  33. Reynolds A, Leake D, Boese Q, Scaringe S, Marshall WS, Khvorova A. Rational siRNA design for RNA interference. Nat Biotechnol. 2004;22:326–30.

    Article  CAS  PubMed  Google Scholar 

  34. Wang J, Place RF, Huang V, Wang X, Noonan EJ, Magyar CE, et al. Prognostic value and function of KLF4 in prostate cancer: RNAa and vector-mediated overexpression identify KLF4 as an inhibitor of tumor cell growth and migration. Cancer Res. 2010;70:10182–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Saito T, Saetrom P. MicroRNAs—targeting and target prediction. New Biotechnol. 2010;27:243–9.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China [grant number 81372759, China]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author information

Author notes

    Authors and Affiliations

    1. Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jie Fang Avenue, Wuhan, 430030, Hubei, China

      Chenghe Wang, Qiangqiang Ge, Zhong Chen, Jia Hu, Fan Li & Zhangqun Ye

    2. Department of Urology, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, No. 600 Yishan Road, Shanghai, 200233, China

      Chenghe Wang

    Authors
    1. Chenghe Wang
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Qiangqiang Ge
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Zhong Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Jia Hu
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Fan Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Zhangqun Ye
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Zhong Chen.

    Ethics declarations

    Conflicts of interest

    None

    Additional information

    Chenghe Wang and Qiangqiang Ge contributed equally to this work.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    ESM 1

    (DOCX 15 kb)

    ESM 2

    (DOCX 14 kb)

    ESM 3

    (DOCX 14 kb)

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Wang, C., Ge, Q., Chen, Z. et al. Promoter-associated endogenous and exogenous small RNAs suppress human bladder cancer cell metastasis by activating p21 CIP1/WAF1 expression. Tumor Biol. 37, 6589–6598 (2016). https://doi.org/10.1007/s13277-015-4571-z

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s13277-015-4571-z

    Keywords

    Search

    Navigation