Real-time quantitative polymerase chain reaction (qPCR) analysis with fluorescence resonance energy transfer (FRET) probes reveals differential expression of the four ERBB4 juxtamembrane region variants between medulloblastoma and pilocytic astrocytoma

N. Zeng, L. Liu, Martin Mccabe, D. T W Jones, K. Ichimura, V. P. Collins

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Aims: We report a comparative study on the mRNA expression of ErbB receptor tyrosine kinases, and in particular ERBB4 transcript variants, in two common paediatric brain tumours: medulloblastoma (MB) and pilocytic astrocytoma (PA). Methods: While the conventional real-time quantitative polymerase chain reaction was used to measure the expression of ERRBs and ErbB4-processing protease genes, the LightCycler fluorescence resonance energy transfer probes were specifically designed to investigate all of the known ERBB4 juxtamembrane (JM) and cytoplasmic transcript variants. Results: The overall expression of ERBBs suggests that ErbB2/ErbB4 heterodimers and ErbB4 homodimers may be major functional units of the ErbBs in MB, while ErbB2/ErbB3 heterodimers may play a more prominent role in addition to ErbB4-containing dimers in PA. Different expression patterns of ERBB4 JM transcripts in MB, PA and normal brain were observed. The JM-d variant was only detected in MBs, while JM-c was present in MB and PA but was not identified in normal brain. The expression of cleavable ERBB4 transcript variants was elevated in PAs and MBs compared with normal brain, while mRNA levels of ErbB4-processing proteases were similar in both tumour types and normal brain. This suggests that proteolytic cleavage of ErbB4 may be more common in MB and PA, which leads to signalling events divergent from those in normal brain. Conclusion: Taken together, these results suggest that ErbB4 processing and function may be altered in brain tumours, such as MB and PA, via differential expression of JM transcript variants. © 2009 Blackwell Publishing Ltd.
    Original languageEnglish
    Pages (from-to)353-366
    Number of pages13
    JournalNeuropathology and Applied Neurobiology
    Volume35
    Issue number4
    DOIs
    Publication statusPublished - Aug 2009

    Keywords

    • ErbB4
    • LightCycler hybridization probe
    • Medulloblastoma
    • Pilocytic astrocytoma
    • Real-time quantitative polymerase chain reaction
    • Transcript variants

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