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

2004; Elsevier BV; Volume: 200; Issue: 7-8 Linguagem: Inglês

10.1016/j.prp.2004.04.008

1618-0631

Torsten Goldmann, Aljoscha M. Flohr, Hugo Murua Escobar, Bernhard Gerstmayer, Uwe Janßen, Andreas Bosio, Siegfried Loeschke, Ekkehard Vollmer, Jörn Bullerdiek,

Cancer-related molecular mechanisms research

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°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.