miRNA Expression Analyses in Prostate Cancer Clinical Tissues.

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).

Journal of visualized experiments : JoVE. 2015 Sep 08*** epublish ***

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

Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco. , Department of Urology, Veterans Affairs Medical Center, San Francisco, University of California San Francisco

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