title: Amplification and overexpression of the IGF2 regulator PLAG1 in hepatoblastoma.
abstract: There is evidence that 8q amplification is associated with poor prognosis in hepatoblastoma. A previous comparative genomic hybridization analysis identified a critical region in chromosomal bands 8q11.2-q13. Using restriction landmark genomic scanning in combination with a virtual genome scan, we showed that this region is delineated by sequences within contig NT_008183 of chromosomal subbands 8q11.22-q11.23. A real-time PCR-based genomic copy number assay of 20 hepatoblastomas revealed gain or amplification in this critical chromosomal region in eight tumors. The expression of four genes and expressed sequence tags (ESTs) within this newly defined region was assayed by real-time reverse transcriptase polymerase chain reaction (RT-PCR) in four tumors with and six tumors without gain or amplification. The PLAG1 oncogene was found to be highly expressed in all but one tumor compared to normal liver tissue. Furthermore, quantitative RT-PCR revealed that the expression level of the developmentally regulated transcription factor PLAG1 was 3-12 times greater in hepatoblastoma tumors and cell lines compared to age-matched normal liver and comparable to the expression in fetal liver tissue. PLAG1 has been shown be a transcriptional activator of IGF2 in other tumor types. Using luciferase reporter assays, we demonstrated that PLAG1 transactivates transcription from the embryonic IGF2 promoter P3, also in hepatoblastoma cell lines. Thus, our results provide evidence that PLAG1 overexpression may be responsible for the frequently observed up-regulation of IGF2 in hepatoblastoma and therefore may be implicated in the molecular pathogenesis of this childhood neoplasia.
authors.affiliation: Institut für Medizinische Biologie, Universität Wien, Vienna, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Zatkova
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Rouillard
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hartmann
authors.forename: Barbara J
authors.initials: BJ
authors.lastname: Lamb
authors.forename: Rork
authors.initials: R
authors.lastname: Kuick
authors.forename: Markus
authors.initials: M
authors.lastname: Eckart
authors.forename: Dietrich
authors.initials: D
authors.lastname: von Schweinitz
authors.forename: Arend
authors.initials: A
authors.lastname: Koch
authors.forename: Christa
authors.initials: C
authors.lastname: Fonatsch
authors.forename: Torsten
authors.initials: T
authors.lastname: Pietsch
authors.forename: Sam M
authors.initials: SM
authors.lastname: Hanash
authors.forename: Katharina
authors.initials: K
authors.lastname: Wimmer
mesh_terms: D000328:Adult; D045744:Cell Line, Tumor; D002648:Child; D002898:Chromosomes, Human, Pair 8 / Q000235:genetics; D019295:Computational Biology / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology; D005333:Fetus; D005784:Gene Amplification / Q000235:genetics*; D018628:Gene Dosage; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D018197:Hepatoblastoma / Q000235:genetics* / Q000473:pathology; D006801:Humans; D007231:Infant, Newborn; D007335:Insulin-Like Growth Factor II / Q000235:genetics*; D008099:Liver / Q000737:chemistry / Q000196:embryology / Q000378:metabolism; D008113:Liver Neoplasms / Q000235:genetics* / Q000473:pathology; D011401:Promoter Regions, Genetic / Q000235:genetics; D015533:Transcriptional Activation / Q000502:physiology; D014162:Transfection
chemical_list: D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; C104382:PLAG1 protein, human; D007335:Insulin-Like Growth Factor II