MET Exon 14 Deletion by PCR
Background: c-Met encodes a receptor tyrosine kinase that is activated by its cognate ligand hepatocyte growth factor (HGF). Phosphorylation of c-Met activates downstream pathways of mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K) and phospholipase C (PLC). Exon 14 is located in the juxtamembrane domain of c-Met and somatic deletion of exon 14 leads to constitutive kinase activation and oncogenesis (Kong-Beltran, et al., 2006). Alternatively spliced variants of Met are found in some gastric and lung carcinomas and may be of prognostic value. In patients with non-small cell lung cancer, MET mutations and amplifications occur in roughly 4-8% and 2-4% respectively.
Methodology: Extracted cellular RNA is run in a real-time, reverse-transcription polymerase chain reaction (RT-PCR) using Taqman® based primer and probe technology. Met Exon 14 Deletions are detected using primer pairs that specifically hybridize the junction sequences flanking Exons 13 and 15. Limit of detection of this assay is 10% mutant total RNA in a background of wild-type RNA.