Summary
DNA cytosine methylation and transcription of specific genes are inversely correlated. In granulocytic differentiation of HL-60 cells there is a distinct down regulation of the c-myc protooncogene expression, which is probably a causal mechanism. With differentiation of HL-60 cells we found no restriction enzyme fragment length polymorphism (RFLP) within the c-myc proto-oncogene, which indicates that there is no loss of regulatory elements (e.g., TATAA boxes within the first exon). Furthermore, we found no de novo methylation in this region. Methylation of other DNA regions, which could influence c-myc expression, is also not necessary for differentiation, as was shown by inhibition of DNA methylase. L-Ethionine and S-adenosyl-L-homocysteine are both potent inhibitors of DNA methylase and do not influence proliferation of HL-60 cells, as shown by FACS analysis.
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References
Alix J-H (1982) Molecular aspects of the in vivo and in vitro effects of ethionine, an analog of methionine. Microbiol Rev, pp 281–295
Bently, DL, Groudin M (1986) A block to elongation is largely responsible for decreased transcription of c-myc in differentiated HL60 cells. Nature 321: 702–706
Blanchard J-M, Piechaczyk M, Dani C, Cahmbard J-C, Franchi A, Pouyssegur J, Jeanteur P (1985) c-myc Gene is transcribed at high rate in GO-arrested fibroblasts and is post-transcriptionally regulated in response to growth factors. Nature 317: 443–435
Bodner AJ, Ting RC, Gallo RC (1981) Induction of differentiation of human promyelocytic leukemia cells (HL-60) by nucleosides and methotrexate. J Natl Cancer Inst 67: 1025–1030
Cheah MSC, Wallace CD, Hoffman RM (1984) Hypomethylation of DNA in human cancer cells: A site-specific change in the c-myc oncogene. J Natl Cancer Inst 73: 1057–1061
Colby WW, Chen EY, Smith DH, Levinson AD (1983) Identification and nucleotide sequence of a human locus homologous to the v-myc oncogene of MC29. Nature 301: 722–725
Collins S, Groudine M (1982) amplification of endogenous myc-related DNA sequences in a human myeloid leukaemia cell line. Nature 298: 679–681
Collins SJ, Ruscetti FW, Gallagher RE, Gallo RC (1978) Terminal differentiation of human promyelocytic leukemia cells induced by dimethyl sulfoxide and other polar compounds. Proc Natl Acad Sci USA 75: 2458–2462
Crooks PA, Tribè MJ, Pinney RJ (1984) Inhibition of bacterial DNA cytosine-5-methyltransferase by S-adenosyl-L-homocysteine and some related compounds. J Pharm Pharmacol 36: 85–89
Dalla Favera R, Wong-Staal F, Gallo RC (1982) Onc Gene amplification in promyelocytic cell line HL-60 and primary leukaemic cells of the same patient. Nature 299: 61–63
Dmitrovsky E, Kuehl M, Hollis GF, Kirsch IR, Bender TP, Segal S (1986) Expression of transfected human c-myc oncogene inhibits differentiation of mouse erythroleukemia cell line. Nature 322: 748–750
Doerfler W (1983) DNA methylation and gene activity. Ann Rev Biochem 52: 93–124
Fearon ER, Winkelstein JA, Civin CJ, Perdall DM, Vogelstein B (1987) Carrier detection in X-linked agammaglobulinemia by analysis of X-chromosomal inactivation. N Engl J Med 316: 427–431
Filmus J, Buick (1985) Relationship of c-myc expression to differentiation proliferation of HL-60 Cells. Cancer Res 45: 822–825
Groffen J, Heisterkamp N, Blennerhassett G, Stevenson JR (1983) Regulation of viral and cellular oncogene expression by cytosine methylation. Virology 126: 213–227
Grosso LE, Pitot HC (1985) Transcriptional regulation of c-myc during chemically induced differentiation of HL-60 cultures. Cancer Res 45: 847–850
Hoffmann RM (1984) Altered methionine metabolism, DNA methylation and oncogene expression in carcinogenesis. Biochim Biophys Acta 738: 49–87
Maniatis T (1984) Molecular cloning. Cold Spring Harbor Laboratory, New York
Resnitsky D, Yarden A, Zipori D, Kimchi A (1986) Autocrine β-related interferon controls c-myc suppression and growth arrest during hematopoetic cell differentiation. Cell 46: 31–40
Shapiro HM (1985) Practical flow cytometry. Liss, New York
Southern EM (1975) Detection of specific DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517
Szyf M, Eliasson L, Mann V, Klein G, Razin A (1985) Cellular and viral DNA hypomethylastion associated with induction of Epstein-Barr virus lytic cycle. Proc Natl. Acad Sci USA 82: 8090–8094
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Supported by the Deutsche Forschungsgemeinschaft (SFB 172, C9) and Sander Stiftung
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Gieseler, F., Meyer, P., Schiffmann, D. et al. Granulocytic differentiation of HL-60 cells is not regulated by DNA de novo methylation. Blut 58, 159–163 (1989). https://doi.org/10.1007/BF00320438
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DOI: https://doi.org/10.1007/BF00320438