Identification of osteopontin as the most consistently over-expressed gene in intrahepatic cholangiocarcinoma: detection by oligonucleotide microarray and real-time PCR analysis.
OBJECTIVE
To investigate the molecular pathways involved in human cholangiocarcinogenesis by gene expression profiling.


METHODS
Oligonucleotide arrays (Affymetrix U133A) were used to establish a specific gene expression profile of intrahepatic CCC in comparison to corresponding non-malignant liver tissue. To validate the expression values of the most overexpressed genes, RT-PCR experiments were performed.


RESULTS
Five hundred and fifty-two statistically differentially expressed genes/ESTs (221 probes significantly up-regulated, 331 probes down-regulated; P < 0.05; fold change > 2; > or = 70%) were identified. Using these data and two-dimensional cluster analysis, a specific gene expression profile was obtained allowing fast and reproducible differentiation of CCC, which was confirmed by supervised neuronal network modelling. The most consistently overexpressed gene (median fold change 33.5, significantly overexpressed in 100%) encoded osteopontin. Furthermore, an association of various genes with the histopathological grading could be demonstrated.


CONCLUSIONS
A highly specific gene expression profile for intrahepatic CCC was identified, allowing for its fast and reproducible discrimination against non-malignant liver tissue and other liver masses. The most overexpressed gene in intrahepatic CCC was the gene encoding osteopontin. These data may lead to a better understanding of human cholangiocarcinogenesis.
Hass
Holger G
HG
Department of Oncology, Hematology and Palliative Care, Marien Hospital, Boeheim-Street 37, Stuttgart 70199, Germany. holgerhass@vinzenz.de
Nehls
Oliver
O
Jobst
Juergen
J
Frilling
Andrea
A
Vogel
Ulrich
U
Kaiser
Stephan
S
D001650:Bile Duct Neoplasms / Q000235:genetics* / Q000601:surgery; D001653:Bile Ducts, Intrahepatic / Q000601:surgery; D018281:Cholangiocarcinoma / Q000235:genetics* / Q000601:surgery; D017931:DNA Primers; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008099:Liver / Q000473:pathology; D008113:Liver Neoplasms / Q000235:genetics* / Q000601:surgery; D020411:Oligonucleotide Array Sequence Analysis; D053495:Osteopontin / Q000235:genetics*; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D053495:Osteopontin