Gene expression array testing of FFPE archival breast tumor samples: an optimized protocol for WG-DASL sample preparation

Breast Cancer Res Treat. 2011 Feb;125(3):879-83. doi: 10.1007/s10549-010-1159-6. Epub 2010 Sep 15.

Abstract

Archived formalin-fixed, paraffin embedded (FFPE) tissues constitute a vast, well-annotated, but underexploited resource for the molecular study of cancer progression, largely because degradation, chemical modification, and cross-linking, render FFPE RNA a suboptimal substrate for conventional analytical methods. We report here a modified protocol for RNA extraction from FFPE tissues which maximized the success rate (with 100% of samples) in the expression profiling of a set of 60 breast cancer samples on the WG-DASL platform; yielding data of sufficient quality such that in hierarchical clustering (a) 12/12 (100%) replicates correctly identified their respective counterparts, with a high self-correlation (r = 0.979), and (b) the overall sample set grouped with high specificity into ER+ (38/40; 95%) and ER- (18/20; 90%) subtypes. These results indicate that a large fraction of decade-old FFPE samples, of diverse institutional origins and processing histories, can yield RNA suitable for gene expression profiling experiments.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Breast / pathology
  • Breast Neoplasms / metabolism*
  • Breast Neoplasms / pathology*
  • Cluster Analysis
  • Cohort Studies
  • Estrogen Receptor alpha / biosynthesis
  • Female
  • Formaldehyde / pharmacology
  • Gene Expression Profiling / methods*
  • Humans
  • Immunohistochemistry / methods
  • Paraffin Embedding / methods
  • RNA, Neoplasm / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Estrogen Receptor alpha
  • RNA, Neoplasm
  • Formaldehyde