| Description: |
The human proto-oncogene C-KIT encodes a receptor tyrosine kinase, and is minimally expressed in normal peripheral blood cells. However, it is commonly expressed in acute myeloid leukemias (AML), in which a spectrum of gain of function mutations has been identified, especially in the core binding factor variants characterized by t(8;21) or inv16(p13q22). Insertions/deletions affecting residue Tyr418 and/or Asp419 in exon 8 lead to hyperactivation. D816V/Y are point mutations affecting exon 17 in the second kinase domain, destabilizing the inactive conformation and hence promoting ligand-independent activation. These two sets of mutations increase the relapse rate and might confer a poor prognosis. C-KIT is expressed in 70%-90% of AML patients, thus the mRNA transcript is a suitable template. The test methodology employs reverse-transcription of the extracted RNA to cDNA. Then, two sets of primers amplify exon 8 and exon 17 using the cDNA as template. The PCR products are being analyzed by direct DNA sequencing. |
