title: Detection of tyrosinase mRNA in formalin-fixed, paraffin-embedded archival sections of melanoma, using the reverse transcriptase in situ polymerase chain reaction.
abstract: Most studies of the reverse transcriptase in situ polymerase chain reaction technique have reported results from assessments of cultured cells, frozen sections, and cytospin preparations. For application to routine diagnosis, it will be necessary to adapt the technique for use with formalin-fixed, paraffin-embedded tissues, the materials that are generally available. We have evaluated the feasibility of such an approach, using surgical pathology archival material from 25 UCLA patients: 15 tissues from primary and metastatic melanoma, 7 from nonmelanocytic tumors, including cancer of the lung, colon, kidney and skin and a thyroid adenoma, and 3 nontumorous tissues. Seven of 15 melanoma tissues gave a strong positive signal, 5 gave a weak signal, and 3 were negative. None of the 10 nonmelanoma tissues gave a positive signal. The specific reaction product was mainly located in the cytoplasm. None of the nonmelanocytic tumors or normal tissues demonstrated this pattern of cytoplasmic staining. Some nonspecific nuclear staining was observed in melanocytic and nonmelanocytic tumors and must not be overread as a true positive result. It is possible to detect tyrosinase mRNA in formalin-fixed, paraffin-embedded tissue sections of melanoma, but the technique remains too demanding for routine application.
authors.affiliation: Department of Pathology, Jonsson Comprehensive Cancer Center, University of California, Los Angeles School of Medicine, California, USA.
authors.forename: J
authors.initials: J
authors.lastname: Guo
authors.forename: L
authors.initials: L
authors.lastname: Cheng
authors.forename: D R
authors.initials: DR
authors.lastname: Wen
authors.forename: R R
authors.initials: RR
authors.lastname: Huang
authors.forename: A J
authors.initials: AJ
authors.lastname: Cochran
mesh_terms: D005557:Formaldehyde; D006801:Humans; D008545:Melanoma / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D014442:Monophenol Monooxygenase / Q000235:genetics*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase; D012878:Skin Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D016707:Tissue Fixation
chemical_list: D012333:RNA, Messenger; D005557:Formaldehyde; D014442:Monophenol Monooxygenase; D012194:RNA-Directed DNA Polymerase