The effect of 50 Hz magnetic field on GCSmRNA expression in lymphoma B cell by mRNA differential display

• Published in: Journal of cellular biochemistry Vol. 79; no. 3; pp. 460 - 470
• Main Authors: Wu, R. Y., Chiang, H., Hu, G. L., Zeng, Q. L., Bao, J. L.
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 01.12.2000
• Subjects: 50 Hz magnetic field; Amino Acid Sequence; Base Sequence; Blotting, Northern; Burkitt Lymphoma - genetics; Burkitt Lymphoma - pathology; Cloning, Molecular; differential display; DNA, Complementary - genetics; Enzyme Induction - radiation effects; GCS mRNA transcription; Gene Expression Profiling; Gene Expression Regulation, Neoplastic - radiation effects; Glucosyltransferases - biosynthesis; Glucosyltransferases - genetics; Humans; Magnetics; Molecular Sequence Data; Neoplasm Proteins - biosynthesis; Neoplasm Proteins - genetics; Polymerase Chain Reaction; RNA, Messenger - biosynthesis; RNA, Neoplasm - biosynthesis; Sequence Homology, Nucleic Acid; Subtraction Technique; Transcription, Genetic - radiation effects; Tumor Cells, Cultured - metabolism; Tumor Cells, Cultured - radiation effects
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/1097-4644(20001201)79:3<460::AID-JCB110>3.0.CO;2-T

Magnetic fields (MFs) of various characteristics can lead to plethora effects in biological system. From a molecular point of view, we hypothesized that there must be a fundamental difference in gene expression between the MF exposed and the unexposed cell. To identify the classes of genes that are regulated, 0.8 mT 50 Hz MF‐induced changes in gene expression were examined in a Daudi cell culture using differential display and reverse transcriptase‐polymerase chain reaction. A candidate cDNA (signatured as MF‐CB) that was observed in the sham‐exposed but not in MF‐exposed cultures was recovered and reamplified. After verification by Northern blot, the cDNA was cloned and sequenced. It was found that 254‐base pair of 5′‐end MF‐CB cDNA clone was identical to gcs in open reading frame (ORF) range. Based on the preliminarily sequence, the prolonged length of 5′‐end MF‐CB cDNA was obtained by PCR amplification and its sequence analysis showed the same results as its original fragment. In order to further determine whether MF‐CB cDNA is from gcs, two Northern blots were probed with gcs and MF‐CB cDNA, respectively, and the data revealed signals of the same size and expression pattern on the two probe filters, which demonstrated that MF‐CB is an EST (expression sequence tag) of gcs. gcs is a gene, identified recently (GenBank accession number D89866), encoding ceramide glucosyltransferase (GCS), which has been implicated as a causal element in human cell growth and differentiation. In an additional experiment, time‐dependent changes in the transcription of gcs induced by 0.8 mT MF were observed by Northern blot with a sharp and reproducible inhibition effect after 20 min exposure and a reduction after 20–24 h exposure. The study demonstrates for the first time that 50 Hz MF can lead to changes in gcs transcription, which provides a new clue to elucidate the mechanism by which MF influence cell growth and differentiation. J. Cell. Biochem. 79:460–470, 2000. © 2000 Wiley‐Liss, Inc.