Detection of HuD transcripts by means of reverse transcriptase and polymerase chain reaction: implications for the detection of minimal residual disease in patients with small cell lung cancer.
Small cell lung cancer (SCLC) expresses neuroectodermal markers including HuD, the best characterized member of the Hu gene family. The aim of this study is to optimize a simple and sensitive reverse transcriptase-polymerase chain reaction assay to detect circulating HuD-expressing cells for the early detection of SCLC recurrences. HuD-specific primers that selectively amplify the three HuD isoforms allowed the detection of one tumor cell/10(6) non-tumor cells. However, HuD transcripts were also detected in the mononuclear fraction of all samples from normal individuals (n=6) and patients with SCLC (n=5). By contrast, HuD protein was not detected in these fractions using Western blotting. More quantitative assays are necessary to examine the value of HuD transcript detection for the identification of tumor recurrences.
Torà
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Unitat de Biologia Cellular i Molecular, Institut Municipal d'Investigació Mèdica, Universitat Autònoma de Barcelona and Universitat Pompeu Fabra, Barcelona, Spain.
Barberá
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Real
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D017398:Alternative Splicing; D000595:Amino Acid Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D018288:Carcinoma, Small Cell / Q000235:genetics / Q000378:metabolism; D016022:Case-Control Studies; D003001:Cloning, Molecular; D003106:Colon / Q000378:metabolism; D003110:Colonic Neoplasms / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D051959:ELAV Proteins; D000067779:ELAV-Like Protein 4; D006801:Humans; D007150:Immunohistochemistry; D008175:Lung Neoplasms / Q000378:metabolism; D008214:Lymphocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D009447:Neuroblastoma / Q000378:metabolism; D010179:Pancreas / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D018076:DNA, Complementary; D051959:ELAV Proteins; D000067779:ELAV-Like Protein 4; C498010:ELAVL4 protein, human; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D016601:RNA-Binding Proteins