Skip to main content

microRNAs in Cancer Chemoprevention: Method to Isolate Them from Fresh Tissues

  • Protocol
Cancer Chemoprevention

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1379))

Abstract

microRNAs are 22-nucleotide-long double-strand small RNAs, able to modulate gene expression at posttranscriptional level, degrading mRNA and/or impairing translation. They have been shown to regulate mRNA and protein abundance and to participate in many regulatory circuits controlling developmental timing, cell proliferation and differentiation, apoptosis and stress response. Notably, microRNA activity has been correlated to the pathogenesis of cancer; they are aberrantly expressed in solid and hematological tumors, suggesting that they could function as oncogenes or tumor suppressors. The emerging role of miRNAs in the carcinogenesis and tumor progression has provided opportunities for their clinical application in the capacity of cancer detection, diagnosis, and prognosis prediction. Here, we describe the experimental protocol used to isolate microRNAs from human tissues coming from head and neck, mesothelioma, and thymoma tumors in order to perform microarray and RT-qPCR experiments.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Lagos-Quintana M, Rauhut R, Lendeckel W et al (2001) Identification of novel genes coding for small expressed RNAs. Science 294:853–858

    Article  CAS  PubMed  Google Scholar 

  2. Tang G, Tang X, Mendu V et al (2008) The art of microRNA: various strategies leading to gene silencing via an ancient pathway. Biochim Biophys Acta 1779:655–662

    Article  CAS  PubMed  Google Scholar 

  3. Ambros V (2004) The functions of animal microRNAs. Nature 431:350–355

    Article  CAS  PubMed  Google Scholar 

  4. Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297

    Article  CAS  PubMed  Google Scholar 

  5. Friedman RC, Farh KK, Burge CB et al (2009) Most mammalian mRNAs are conserved targets of microRNAs. Genome Res 19:92–105

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Landgraf P, Rusu M, Sheridan R et al (2007) A mammalian microRNA expression atlas based on small RNA library sequencing. Cell 129:1401–1414

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Ross JS, Carlson JA, Brock G (2007) miRNA: the new gene silencer. Am J Clin Pathol 128:830–836

    Article  CAS  PubMed  Google Scholar 

  8. Visone R, Croce CM (2009) MiRNAs and cancer. Am J Pathol 174:1131–1138

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Lu J, Getz G, Miska EA et al (2005) MicroRNA expression profiles classify human cancers. Nature 435:834–838

    Article  CAS  PubMed  Google Scholar 

  10. Pritchard CC, Cheng HH, Tewari M (2012) MicroRNA profiling: approaches and considerations. Nat Rev Genet 13:358–369

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Li J, Smyth P, Flavin R et al (2007) Comparison of miRNA expression patterns using total RNA extracted from matched samples of formalin-fixed paraffin-embedded (FFPE) cells and snap frozen cells. BMC Biotechnol 7:36

    Article  PubMed Central  PubMed  Google Scholar 

  12. Cioce M, Ganci F, Canu V et al (2014) Protumorigenic effects of mir-145 loss in malignant pleural mesothelioma. Oncogene 33:5319

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Ganci F, Sacconi A, Bossel Ben-Moshe N et al (2013) Expression of TP53 mutation-associated microRNAs predicts clinical outcome in head and neck squamous cell carcinoma patients. Ann Oncol 24:3082–3088

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Ganci F, Vico C, Korita E et al (2014) MicroRNA expression profiling of thymic epithelial tumors. Lung Cancer 85:197–204

    Article  PubMed  Google Scholar 

  15. Mraz M, Malinova K, Mayer J et al (2009) MicroRNA isolation and stability in stored RNA samples. Biochem Biophys Res Commun 390:1–4

    Article  CAS  PubMed  Google Scholar 

  16. Podolska A, Kaczkowski B, Litman T et al (2011) How the RNA isolation method can affect microRNA microarray results. Acta Biochim Pol 58:535–540

    CAS  PubMed  Google Scholar 

  17. Hammerle-Fickinger A, Riedmaier I, Becker C et al (2010) Validation of extraction methods for total RNA and miRNA from bovine blood prior to quantitative gene expression analyses. Biotechnol Lett 32:35–44

    Article  CAS  PubMed  Google Scholar 

  18. Ibberson D, Benes V, Muckenthaler MU et al (2009) RNA degradation compromises the reliability of microRNA expression profiling. BMC Biotechnol 9:102

    Article  PubMed Central  PubMed  Google Scholar 

  19. Wang WX, Wilfred BR, Baldwin DA et al (2008) Focus on RNA isolation: obtaining RNA for microRNA (miRNA) expression profiling analyses of neural tissue. Biochim Biophys Acta 1779:749–757

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. Liu A, Tetzlaff MT, Vanbelle P et al (2009) MicroRNA expression profiling outperforms mRNA expression profiling in formalin-fixed paraffin-embedded tissues. Int J Clin Exp Pathol 2:519–527

    PubMed Central  CAS  PubMed  Google Scholar 

  21. Zhang X, Chen J, Radcliffe T et al (2008) An array-based analysis of microRNA expression comparing matched frozen and formalin-fixed paraffin-embedded human tissue samples. J Mol Diagn 10:513–519

    Article  PubMed Central  PubMed  Google Scholar 

  22. Doleshal M, Magotra AA, Choudhury B et al (2008) Evaluation and validation of total RNA extraction methods for microRNA expression analyses in formalin-fixed, paraffin-embedded tissues. J Mol Diagn 10:203–211

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Giovanni Blandino .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Ganci, F., Blandino, G. (2016). microRNAs in Cancer Chemoprevention: Method to Isolate Them from Fresh Tissues. In: Strano, S. (eds) Cancer Chemoprevention. Methods in Molecular Biology, vol 1379. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3191-0_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3191-0_3

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3190-3

  • Online ISBN: 978-1-4939-3191-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics