Reliable microRNA profiling in routinely processed formalin-fixed paraffin-embedded breast cancer specimens using fluorescence labelled bead technology

BMC Biotechnol. 2008 Nov 27:8:90. doi: 10.1186/1472-6750-8-90.

Abstract

Background: During the last years the analysis of microRNA expression patterns has led to completely new insights into cancer biology. Furthermore, these patterns are a very promising tool for the development of new diagnostic and prognostic markers. However, most human tumour samples for which long term clinical records are available exist only as formalin-fixed paraffin-embedded specimens. Therefore, the aim of this study was to examine the feasibility of microRNA profiling studies in routinely processed formalin-fixed paraffin-embedded human breast cancer specimens using fluorescence labelled bead technology.

Results: A statistically highly significant correlation (Spearman r: 0.78 - 0.90, p < 0.0001) was observed for the expression of 319 microRNAs in routinely processed FFPE breast cancer specimens and paired fresh frozen tissue samples (n = 5). Results were confirmed in a larger series analyzing a selection of 10 microRNAs reported to be deregulated in breast cancer (n = 12). The expression pattern of 3 microRNAs was independently validated in this cohort using real-time RT-PCR technology.

Conclusion: Comprehensive microRNA expression patterns can be reliably derived from routinely processed FFPE breast cancer specimens using fluorescence labelled bead technology.

MeSH terms

  • Breast Neoplasms / genetics*
  • Fluorescent Dyes / metabolism*
  • Formaldehyde / metabolism
  • Frozen Sections
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Neoplastic
  • Humans
  • MicroRNAs / genetics*
  • MicroRNAs / standards
  • Microspheres
  • Paraffin Embedding / methods*
  • Quality Control
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Staining and Labeling / methods*
  • Tissue Fixation / methods*

Substances

  • Fluorescent Dyes
  • MicroRNAs
  • Formaldehyde