Development of a single-step duplex RT-PCR detecting different forms of ret activation, and identification of the third form of in vivo ret activation in human papillary thyroid carcinoma.
In up to 25% of human papillary thyroid carcinomas (PCs), the oncogenic activation of ret results from the fusion of its tyrosine kinase domain with different unlinked amino-terminal sequences. Activation of this oncogene may be of prognostic significance in patients with PC. To screen for ret activation in archival paraffin-embedded tumors, we have developed a single-step duplex RT-PCR to detect different forms of ret activation despite different chimeric transcripts being expressed. Furthermore, we report a third type of ret oncogenic activation, named ret/PTC3, identified by using this novel method followed by rapid amplification of cDNA ends (5'-RACE) cloning.
Jhiang
S M
SM
Department of Physiology, Ohio State University, Columbus 43210.
Smanik
P A
PA
Mazzaferri
E L
EL
D000595:Amino Acid Sequence; D001483:Base Sequence; D002277:Carcinoma / Q000235:genetics*; D029721:Drosophila Proteins; D015972:Gene Expression Regulation, Neoplastic; D015321:Gene Rearrangement; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D009857:Oncogenes; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051096:Proto-Oncogene Proteins c-ret; D012334:RNA, Neoplasm / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D013964:Thyroid Neoplasms / Q000235:genetics*
D029721:Drosophila Proteins; D011518:Proto-Oncogene Proteins; D012334:RNA, Neoplasm; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases; C416649:Ret protein, Drosophila