title: Microarray analysis reveals differential expression of benign and malignant pheochromocytoma.
abstract: The diagnosis of a malignant pheochromocytoma (PC) can only be established by the presence of distant metastases, but a subset of apparently benign PCs develop metastases. We have employed a microarray analysis to identify a typical gene expression profile which distinguishes malignant from benign PC. Total RNA was isolated from fresh-frozen tissue of five benign and five malignant PCs. The reference consisted of laser microdissected tissue from normal adrenal medulla. After generating Cy3- and Cy5-fluorescently labeled cDNAs, F-chips containing 11 540 spots were hybridized. Data were analyzed with the IMAGENE 3.0 software. Gene expression levels were validated by real-time (RT)-PCR and immunohistochemistry (IHC). The analysis revealed a more than twofold difference in expression between benign and malignant PCs in 132 genes: 19 were up-regulated and 113 were down-regulated. Expression differences of six genes (calsequestrin, NNAT, neurogranin, secreted protein acidic and rich in cysteine (SPARC), EGR2, and MAOB) were confirmed by RT-PCR in 25 PCs. IHC for calsequestrin revealed an overexpression in malignant PCs (7/10 vs 1/10, P=0.03). Comparative analysis by microarray of all ten PCs (benign/malignant) versus normal adrenal medulla revealed a more than twofold expression difference in 455/539 and 491/671 genes respectively. Several of these genes are known to participate on adrenal tumorigenesis, potential tumor suppressor genes, and oncogenes. Comprehensive gene expression analysis of malignant and benign PCs revealed different gene profiles, which could be used to discriminate between malignant and benign PCs. Based on these findings, the strategy for further follow-up and treatment could be modified accordingly.
authors.affiliation: Department of Surgery, Philipps-Universität Marburg, Baldingerstrasse, D-35043 Marburg, Germany. jwaldman@med.uni-marburg.de
authors.forename: Jens
authors.initials: J
authors.lastname: Waldmann
authors.forename: Volker
authors.initials: V
authors.lastname: Fendrich
authors.forename: Julia
authors.initials: J
authors.lastname: Holler
authors.forename: Malte
authors.initials: M
authors.lastname: Buchholz
authors.forename: Ernst
authors.initials: E
authors.lastname: Heinmöller
authors.forename: Peter
authors.initials: P
authors.lastname: Langer
authors.forename: Annette
authors.initials: A
authors.lastname: Ramaswamy
authors.forename: Birgit
authors.initials: B
authors.lastname: Samans
authors.forename: Martin K
authors.initials: MK
authors.lastname: Walz
authors.forename: Matthias
authors.initials: M
authors.lastname: Rothmund
authors.forename: Detlef K
authors.initials: DK
authors.lastname: Bartsch
authors.forename: Emily P
authors.initials: EP
authors.lastname: Slater
mesh_terms: D000310:Adrenal Gland Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000313:Adrenal Medulla / Q000378:metabolism* / Q000473:pathology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D016022:Case-Control Studies; D020869:Gene Expression Profiling; D006801:Humans; D007124:Immunoenzyme Techniques; D008875:Middle Aged; D009362:Neoplasm Metastasis; D020411:Oligonucleotide Array Sequence Analysis; D010673:Pheochromocytoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055815:Young Adult
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger