title: Development of a robust protocol for gene expression analysis using formalin-fixed, paraffin-embedded liver transplant biopsy specimens.
abstract: Liver transplant biopsies are routinely archived following formalin fixation and paraffin embedding and may provide an additional source of diagnostic information following transcriptomic biomarker analysis. This study was designed to compare gene transcription between resting and stressed biliary cells in culture, these cells after fixation and embedding and archival liver transplant biopsy tissue. The transcription of p21/WAF1 and transforming growth factor (TGF)-β1 showed similar changes in the fresh and embedded liver cells. However, the expression of TGF-β2 was markedly different between the fresh and embedded samples, suggesting that fixation can produce sequence-specific artefacts. Sufficient quantities of pure RNA were recovered from all the liver transplant biopsies to allow complementary DNA production. Measurement of the transcription of all three genes showed variability between the cases. Although the results for individual transcripts should be interpreted with care, these data do suggest the feasibility of performing a larger biomarker discovery studies using archival tissue.
authors.affiliation: Applied Immunobiology and Transplantation Research Group, Faculty of Medical Sciences, Institute of Cellular Medicine, Medical School, Newcastle University, Newcastle upon Tyne, Tyne and Wear, UK.
authors.forename: Emily
authors.initials: E
authors.lastname: Thompson
authors.forename: Alastair D
authors.initials: AD
authors.lastname: Burt
authors.forename: Catriona E
authors.initials: CE
authors.lastname: Barker
authors.forename: John A
authors.initials: JA
authors.lastname: Kirby
authors.forename: John G
authors.initials: JG
authors.lastname: Brain
mesh_terms: D005557:Formaldehyde; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D016031:Liver Transplantation; D016612:Paraffin Embedding; D016707:Tissue Fixation / Q000379:methods
chemical_list: D005557:Formaldehyde
keywords: Biliary; Liver; Transplantation