MicroRNA expression profiling of specific cells in complex archival tissue stained by immunohistochemistry.

Author(s): Schuster C, Budczies J, Faber C, Kirchner T, Hlubek F

Publication: Lab Invest, 2011, Vol. 91, Page 157-65

PubMed ID: 20661226 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of staining with IHC or hemalaun, and IHC antigen retrieval temperature on RNA integrity and miRNA expression in FFPE colorectal carcinoma specimens.

   Summary of Findings:

   Reducing the antigen retrieval temperature from 98 to 90 degrees C caused a slight reduction in B-catenin nuclear staining. The authors report that when the antigen retrieval temperature was reduced to 80 degrees C, there was no staining for B-catenin. Regardless of the antigen retrieval temperature, capillary electrophoresis showed complete loss of the 18S and 28S ribosomal RNA peaks in all IHC specimens and in hemalaun-stained specimens which were not exposed to elevated temperatures. Hemalaun-stained and IHC specimens had a peak of RNA at 130 bp, but the peak was more pronounced in hemalaun-stained specimens. Despite exposure to heat, the relative abundance of 5 different miRNAs did not differ between IHC and hemalaun-stained specimens. Using laser-microdissected beta-catenin expressing cells, distinct but reproducible patterns of miRNA expression were observed between patients. The authors conclude that miRNA expression profiling can be performed on IHC or pathology stained archived FFPE tumor specimens.

   Biospecimens

   • Tissue - Colorectal

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry
   Morphology	Light microscopy
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Immunohistochemistry Specific	Targeted peptide/protein	B-catenin
   Biospecimen Acquisition	Pathology ink	Hemalaun IHC stained
   Real-time qRT-PCR Specific	Targeted nucleic acid	miR16 miR206 miR214 miR197 miR191 miR19b miR7 miR181 miR195 miR196b
   Analyte Extraction and Purification	Temperature of heat-induced retrieval	98 degrees C 90 degrees C 80 degrees C

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