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


CONCLUSIONS
Comprehensive microRNA expression patterns can be reliably derived from routinely processed FFPE breast cancer specimens using fluorescence labelled bead technology.
Hasemeier
Britta
B
Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany. Hasemeier.Britta@MH-Hannover.de
Christgen
Matthias
M
Kreipe
Hans
H
Lehmann
Ulrich
U
D001943:Breast Neoplasms / Q000235:genetics*; D005456:Fluorescent Dyes / Q000378:metabolism*; D005557:Formaldehyde / Q000378:metabolism; D005629:Frozen Sections; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000592:standards; D008863:Microspheres; D016612:Paraffin Embedding / Q000379:methods*; D011786:Quality Control; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013194:Staining and Labeling / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods*
D005456:Fluorescent Dyes; D035683:MicroRNAs; D005557:Formaldehyde