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

• Published in: Cancer letters 2000-12, Vol.161 (2), p.157-164
• Main Authors: Torà, Montserrat, Barberá, Vı́ctor M, Real, F.X
• Format: Article
• Language: English
• Subjects: Alternative Splicing; Amino Acid Sequence; Biological and medical sciences; Blotting, Southern; Blotting, Western; Carcinoma, Small Cell - genetics; Carcinoma, Small Cell - metabolism; Case-Control Studies; Cloning, Molecular; Colon - metabolism; Colonic Neoplasms - metabolism; DNA Primers; DNA, Complementary - metabolism; ELAV Proteins; ELAV-Like Protein 4; HuD; Humans; Immunohistochemistry; Lung Neoplasms - metabolism; Lymphocytes - metabolism; Medical sciences; Minimal residual disease; Molecular Sequence Data; Nerve Tissue Proteins - biosynthesis; Neuroblastoma; Neuroblastoma - metabolism; Pancreas - metabolism; Pancreatic Neoplasms - metabolism; Pneumology; Polymerase Chain Reaction; Protein Isoforms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; RNA-Binding Proteins - biosynthesis; Sequence Homology, Amino Acid; Small cell lung cancer; Tumor Cells, Cultured; Tumors of the respiratory system and mediastinum
• ISSN: 0304-3835; 1872-7980
• DOI: 10.1016/S0304-3835(00)00577-2

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.