The HOPE-technique permits Northern blot and microarray analyses in paraffin-embedded tissues

Pathol Res Pract. 2004;200(7-8):511-5. doi: 10.1016/j.prp.2004.04.008.

Abstract

There is an increasing demand for tissue samples that, after having been used for conventional histologic examination, are also suited for molecular analyses. As to formalin-fixed, paraffin embedded (FFPE) tissue, the latter applications are very limited. The HOPE (Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) technique comprises a new protection-solution with an organic buffer, with acetone as the only dehydrating agent, and pure paraffin of 52-54 degrees C melting temperature, allowing for all pathologic routine investigations. In contrast to FFPE tissue, the HOPE-technique allows for the application of molecular methods, such as high molecular DNA and RNA isolation, which can be used for PCR and reverse transcription PCR (RT-PCR). In this study, we investigated whether RNA from HOPE-fixed tissue samples is suitable for Northern blot and microarray analyses. RNAs of two HOPE-fixed breast cancer specimens of different histologic grade were used to carry out an array experiment. It turned out that RNA from HOPE-fixed tissue is of high quality and can be successfully used for array experiments. In addition, by detecting GAPDH and high mobility group protein gene B1 (HMGB1)-specific transcripts, we were able to demonstrate that RNA from HOPE-fixed tissue can also be used for Northern blot hybridization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Northern*
  • Breast Neoplasms / genetics*
  • Female
  • Glutamic Acid / pharmacology
  • Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
  • HEPES / pharmacology
  • HMGB1 Protein / genetics
  • Humans
  • Oligonucleotide Array Sequence Analysis*
  • RNA / metabolism*
  • Solvents / pharmacology
  • Tissue Fixation / methods*

Substances

  • HMGB1 Protein
  • Solvents
  • Glutamic Acid
  • RNA
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • HEPES