Keywords
RET gene mutations
next-generation sequencing
How to Cite
Abstract
Abstract. Medullary thyroid carcinoma (MTC) arises from non-epithelial parafollicular cells. MTC can be hereditary (25%) or sporadic (75%). Activating germline REarranged during Transfection (RET) mutations are found in 95-98% of hereditary MTC, while somatic RET mutations are present in 25-40% of sporadic MTC.
Aim. The aim of the study was to determine RET gene mutations in codons 634 and 918, which are associated with the highest risk of MTC.
Material and methods. Whole blood was obtained by standard venipuncture. Genomic DNA was isolated using the EliGene® Blood DNA Isolation kit. DNA amplification was performed by polymerase chain reaction (PCR) using primers specific for the regions of exons 10, 11 and 16 of the RET gene, which may contain mutations. The next-generation sequencing (NGS) for genomic DNAs from each sample was performed using the Ion Ampliseq Cancer Hotspot Panel v2 following the manufacturer's instructions.
Results. Among all the tested samples, heterozygous p.M918T mutation was detected only in sample No. 2. As an outcome of the sequencing of the genomic DNA of 7 patients diagnosed with MTC, four different pathogenic inherited mutations of the RET gene were identified in four patients.
Conclusions. Determination of mutations in the RET gene is a promising approach for establishing a diagnosis in the treatment of hereditary forms of MTC and allows planning further directions of therapy. The PCR-restriction fragment length polymorphism (PCR-RFLP) method makes it possible to detect individual mutations in the RET and other genes, associated with the highest risk of MTC (M918T, C634F and A883F). Given the population frequencies of occurrence of various pathogenic variants in the RET gene in Ukraine, the (PCR-RFLP) method can become an effective screening tool. NGS allows analyzing the entire sequence of the RET gene and other genes, and detecting a much larger spectrum of variants, including previously undescribed ones.
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