1. Comparative analysis of expression of mutant and wild-type alleles is essential for reliable PCR-based detection of MET exon 14 skipping
- Author
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Alexandr O. Ivantsov, Natalia V. Mitiushkina, Tatiana N. Sokolova, Maxim M. Kholmatov, Ilya A. Stepanov, Vladislav I. Tiurin, Ekatherina Sh. Kuligina, Olga S. Yatsuk, Alexandr A. Romanko, Alexandr V. Togo, Alexey M. Belyaev, and Evgeny N. Imyanitov
- Subjects
Adult ,Male ,0301 basic medicine ,Lung Neoplasms ,MET Exon 14 Mutation ,Adenocarcinoma of Lung ,Biology ,Biochemistry ,Young Adult ,03 medical and health sciences ,Exon ,symbols.namesake ,Carcinoma, Non-Small-Cell Lung ,Humans ,Allele ,Gene ,Alleles ,Aged ,Aged, 80 and over ,Sanger sequencing ,Splice site mutation ,030102 biochemistry & molecular biology ,Alternative splicing ,Wild type ,Exons ,General Medicine ,Middle Aged ,Proto-Oncogene Proteins c-met ,Molecular biology ,030104 developmental biology ,Mutation ,symbols ,Female - Abstract
MET exon 14 skipping (exon 14Δ) mutations are associated with tumor sensitivity to a number of tyrosine kinase inhibitors, however clinical testing for MET gene status remains complicated. We developed a simple allele-specific PCR cDNA-based test, which allowed for the identification of MET exon 14Δ allele in 35 (2.5%) out of 1415 EGFR mutation–negative lung carcinomas (LCs). MET exon 14Δ was significantly associated with elderly age and non-smoking status of the patients. A total of 34 (97%) out of 35 tumors carrying MET exon 14Δ showed preferential expression of the mutated allele; this imbalance was attributed to the down-regulation of the expression of the wild-type gene copy. Sanger sequencing confirmed the presence of genomic exon 14 splice site mutations in 24/35 (68.6%) cases, which showed MET exon 14 skipping by PCR. In addition to LCs described above, some carcinomas demonstrated low-abundance MET exon 14Δ-specific signal. Low-level expression of MET exon 14Δ allele may potentially compromise the results of allele-specific PCR-based tests, therefore comparison of the level of expression of mutated and normal alleles is essential for the reliability of MET gene testing.
- Published
- 2019
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