1. Quantification ofPDGFRAalternative transcripts improves the screening forX–PDGFRAfusions by reverse transcriptase-polymerase chain reaction
- Author
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José L. Vizmanos, María José Calasanz, M. García-Delgado, C. Hurtado, I. Erquiaga, Francisco J. Novo, Paula Aranaz, and Cristina Ormazábal
- Subjects
Male ,Cancer Research ,Receptor, Platelet-Derived Growth Factor alpha ,Oncogene Proteins, Fusion ,Oncogene Proteins ,PDGFRB ,PDGFRA ,Biology ,law.invention ,Fusion gene ,law ,Cell Line, Tumor ,Eosinophilia ,Humans ,RNA, Messenger ,Gene ,In Situ Hybridization, Fluorescence ,Polymerase chain reaction ,Regulation of gene expression ,Reverse Transcriptase Polymerase Chain Reaction ,Hematology ,Prognosis ,Molecular biology ,digestive system diseases ,Reverse transcriptase ,Gene Expression Regulation, Neoplastic ,Alternative Splicing ,Oncology ,Case-Control Studies ,Hematologic Neoplasms ,Cancer research ,Female - Abstract
Hematological malignancies with eosinophilia are often associated with fusions in PDGFRA, PDGFRB, or FGFR1 genes. RT-PCR has proved to be useful for finding new PDGFRA gene fusions, but some studies have shown overexpression of the TK domain which cannot be explained by the existence of such aberrations. This fact could be related to the expression of alternative PDGFRA transcripts. We show that quantification of the expression of three different PDGFRA fragments discriminates between PDGFRA alternative transcripts and fusion genes, and we have tested this novel methodological approach in a group of eosinophilia cases. Our data show that alternative PDGFRA transcripts should be taken into account when screening for PDGFRA aberrations, such as gene fusions, by RT-PCR. Expression from an internal PDGFRA promoter seems to be a frequent event, in both normal and leukemic samples, and is probably related to physiological conditions, but it could have a role in other tumors. Even so, we show that our RQ-PCR methodology can discriminate expression of alternative transcripts from the presence of X-PDGFRA fusion genes.
- Published
- 2010