miRNA expression analyses in prostate cancer clinical tissues

Nathan Bucay, Varahram Shahryari, Shahana Majid, Soichiro Yamamura, Yozo Mitsui, Z. Laura Tabatabai, Kirsten Greene, Guoren Deng, Rajvir Dahiya, Yuichiro Tanaka, Sharanjot Saini

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

A critical challenge in prostate cancer (PCa) clinical management is posed by the inadequacy of currently used biomarkers for disease screening, diagnosis, prognosis and treatment. In recent years, microRNAs (miRNAs) have emerged as promising alternate biomarkers for prostate cancer diagnosis and prognosis. However, the development of miRNAs as effective biomarkers for prostate cancer heavily relies on their accurate detection in clinical tissues. miRNA analyses in prostate cancer clinical specimens is often challenging owing to tumor heterogeneity, sampling errors, stromal contamination etc. The goal of this article is to describe a simplified workflow for miRNA analyses in archived FFPE or fresh frozen prostate cancer clinical specimens using a combination of quantitative real-time PCR (RT-PCR) and in situ hybridization (ISH). Within this workflow, we optimize the existing methodologies for miRNA extraction from FFPE and frozen prostate tissues and expression analyses by Taqman-probe based miRNA RT-PCR. In addition, we describe an optimized method for ISH analyses formiRNA detection in prostate tissues using locked nucleic acid (LNA)- based probes. Our optimized miRNA ISH protocol can be applied to prostate cancer tissue slides or prostate cancer tissue microarrays (TMA).

Original languageEnglish (US)
Article numbere53123
JournalJournal of Visualized Experiments
Volume2015
Issue number103
DOIs
StatePublished - Sep 8 2015
Externally publishedYes

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Keywords

  • Clinical tissues
  • Expression analyses
  • In situ hybridization
  • Issue 103
  • Medicine
  • MicroRNAs
  • Prostate cancer
  • Real-time PCR

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Bucay, N., Shahryari, V., Majid, S., Yamamura, S., Mitsui, Y., Tabatabai, Z. L., Greene, K., Deng, G., Dahiya, R., Tanaka, Y., & Saini, S. (2015). miRNA expression analyses in prostate cancer clinical tissues. Journal of Visualized Experiments, 2015(103), [e53123]. https://doi.org/10.3791/53123