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Updated: May 10, 2026
Quantitative Analysis of Alternative Pre-mRNA Splicing in Mouse Brain Sections Using RNA In Situ Hybridization Assay
Published on: August 26, 2018
Identification of MET genomic amplification, protein expression and alternative splice isoforms in neuroblastomas
Benedict Yan1, Malcolm Lim, Lihan Zhou
1Department of Pathology and Laboratory Medicine, KK Women's and Children's Hospital, Singapore.
Background:
Crizotinib, a dual anaplastic lymphoma kinase (ALK) and mesenchymal-epithelial transition (MET) tyrosine kinase inhibitor, is currently being evaluated for the treatment of neuroblastoma. Its effects are thought to be mediated mainly via its activity against ALK. Although MET genomic/protein expression status might conceivably affect crizotinib efficacy, this issue has hitherto not received attention in neuroblastomas.
Aims/Methods:
MET genomic and protein expression status was characterised by silver in situ hybridisation and immunohistochemistry (IHC) respectively, in a cohort of 54 neuroblastoma samples. MET splice isoforms were characterised in 15 of these samples by quantitative PCR.
Results:
One case (1/54; prevalence 1.85%) displayed MET genomic amplification, while another case (1/54; prevalence 1.85%) displayed strong membranous MET protein expression (IHC score 3+). Alternative exon 10-deleted and exon 14-deleted MET splice isoforms were identified.
Conclusions:
MET amplification and protein expression, although low in prevalence, are present in neuroblastomas. This has implications when crizotinib is employed as a therapeutic agent in neuroblastomas. Additionally, the existence of alternatively spliced MET isoforms may have clinical and biological implications in neuroblastomas.
Insights
Mesenchymal-epithelial transition (MET) amplification and protein expression occur in neuroblastoma, impacting crizotinib treatment. Alternative MET splice variants also present potential clinical implications for this cancer.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Crizotinib, an anaplastic lymphoma kinase (ALK) and MET inhibitor, is under investigation for neuroblastoma treatment.
- Crizotinib's efficacy is primarily attributed to its ALK inhibition, with MET's role less understood in neuroblastoma.
- The impact of MET genomic and protein expression on crizotinib efficacy in neuroblastoma remains unexplored.
Purpose of the Study:
- To investigate the prevalence of MET genomic amplification and protein expression in neuroblastoma.
- To characterize MET splice isoforms in neuroblastoma samples.
- To assess the potential implications of MET status for crizotinib therapy in neuroblastoma.
Main Methods:
- Genomic DNA and protein were analyzed using silver in situ hybridization and immunohistochemistry (IHC) in 54 neuroblastoma samples.
- Quantitative PCR was employed to identify and characterize MET splice isoforms in 15 neuroblastoma samples.
- The study focused on MET amplification, protein expression levels (IHC score), and alternative splicing.
Main Results:
- MET genomic amplification was detected in 1.85% (1/54) of neuroblastoma cases.
- Strong membranous MET protein expression (IHC score 3+) was observed in 1.85% (1/54) of cases.
- Alternative MET splice isoforms, including exon 10-deleted and exon 14-deleted variants, were identified.
Conclusions:
- MET amplification and protein expression, though infrequent, are present in neuroblastoma.
- These findings suggest potential implications for crizotinib treatment strategies in neuroblastoma.
- The presence of alternatively spliced MET isoforms may hold clinical and biological significance for neuroblastoma patients.
