The HOPE-technique permits Northern blot and microarray analyses in paraffin-embedded tissues.
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.
Clinical and Experimental Pathology, Research Center Borstel, Parkallee 3, 23845 Borstel, Germany.
Torsten
T
Goldmann
Aljoscha M
AM
Flohr
Hugo
H
Murua Escobar
Bernhard
B
Gerstmayer
Uwe
U
Janssen
Andreas
A
Bosio
Siegfried
S
Loeschke
Ekkehard
E
Vollmer
Jörn
J
Bullerdiek
D015152:Blotting, Northern; D001943:Breast Neoplasms / Q000235:genetics*; D005260:Female; D018698:Glutamic Acid / Q000494:pharmacology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006531:HEPES / Q000494:pharmacology; D024243:HMGB1 Protein / Q000235:genetics; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000378:metabolism*; D012997:Solvents / Q000494:pharmacology; D016707:Tissue Fixation / Q000379:methods*
D024243:HMGB1 Protein; D012997:Solvents; D018698:Glutamic Acid; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D006531:HEPES