Mutations and elevated transcriptional activity of conductin (AXIN2) in hepatoblastomas.
Hepatoblastoma (HB) is the most frequent malignant liver tumour of childhood. Most HBs develop sporadically but their incidence is highly elevated in patients with familial adenomatous polyposis coli (FAP). These patients carry germline mutations in the adenomatous polyposis coli (APC) tumour suppressor gene. APC forms a multi-protein complex involved in the WNT signalling pathway that controls the stability of beta-catenin, the central effector in this cascade. Whereas APC mutations are rare in sporadic HBs, a high frequency of beta-catenin mutations leading to overactivation of WNT signalling was previously found in these tumours. This pathway is negatively controlled by conductin (axin2), representing a further partner in this signalling complex. To investigate whether alterations in conductin may also be involved in the pathogenesis of sporadic HBs, 37 HBs and five HB cell lines were screened for mutations using single-strand conformation polymorphism (SSCP) analysis, reverse transcription-polymerase chain reaction (RT-PCR), and direct sequencing. In two cases, larger deletions (52 and 1624 bp) leading to frameshifts were found. In addition, one HB carried a somatic point mutation. Expression analysis by competitive RT-PCR in HBs revealed up-regulation of conductin mRNA compared with adjacent liver samples. This mRNA overexpression resulted in increased conductin protein levels demonstrated by western blot analysis. Tumours with activating beta-catenin mutations revealed higher levels of conductin mRNA transcripts. This finding indicates that conductin is a direct target gene of WNT signalling in HBs, as has been demonstrated in other tissues. In summary, conductin mutations may represent an alternative mechanism leading to activation of WNT signalling in HBs. The overexpression of conductin mRNA in HBs reflects activation of the WNT pathway because conductin represents a target gene of WNT signalling in liver tissue.
Koch
Arend
A
Department of Neuropathology, University of Bonn Medical Center, Bonn, Germany.
Weber
Nina
N
Waha
Andreas
A
Hartmann
Wolfgang
W
Denkhaus
Dorota
D
Behrens
Jürgen
J
Birchmeier
Walter
W
von Schweinitz
Dietrich
D
Pietsch
Torsten
T
D060466:Axin Protein; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D002675:Child, Preschool; D003598:Cytoskeletal Proteins / Q000235:genetics*; D017353:Gene Deletion; D018197:Hepatoblastoma / Q000235:genetics*; D006801:Humans; D007223:Infant; D008099:Liver / Q000378:metabolism; D008113:Liver Neoplasms / Q000235:genetics*; D009154:Mutation; D018807:Polymorphism, Single-Stranded Conformational; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015534:Trans-Activators / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics*; D015854:Up-Regulation / Q000235:genetics; D051176:beta Catenin
C491761:AXIN2 protein, human; D060466:Axin Protein; C495270:CTNNB1 protein, human; D003598:Cytoskeletal Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D015534:Trans-Activators; D051176:beta Catenin