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A novel MASH1 enhancer with N-myc and CREB-binding sites is active in neuroblastoma

Cancer Gene Therapy volume 14, pages 287–296 (2007)Cite this article

Abstract

Neuroblastoma is one of the most common solid tumors in childhood. With the aim of developing a targeting vector for neuroblastoma, we cloned and characterized an enhancer in the 5′-flanking regions of the MASH1 gene by a random-trap method from a 36 kb cosmid DNA. The enhancer-containing clone was identified by the expression of GFP when transfected into neuroblastoma cell lines. The enhancer-luciferase activity is higher in neuroblastoma cell lines, IMR32, BE2 and SH-SY5Y, compared with those in non-neuroblastoma cell lines, U1242 glioma, N417 small cell lung cancer and EOMA hemangioma. The core enhancer was determined within a 0.2 kb fragment, yielding three- to fourfold higher activity than that of the MASH1 promoter alone in IMR32 and BE2. This area possesses GATA- and CREB-binding sites, as well as the E-box. EMSA on this area demonstrated that CREB/ATF could bind the DNA. Chromatin immunoprecipitation assay revealed that N-myc, CREB, and co-activators CBP and PCAF, but not HDAC1, are bound to the core enhancer at the same time as the co-activators and N-myc bind to the promoter. This supports the idea that the commonly overexpressed genes HASH1 and N-myc are regulated in concert, confirming their importance as prognostic markers or targets for therapy.

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Acknowledgements

We are grateful to Dr J Johnson for the McosI cosmid and her valuable comment on the manuscript, as well as to Drs O Nilsson, S Påhlman, RA Reisfeld, L Welsh, and B Westermark for cell lines. We thank K Björklund and T Olsson for excellent technical assistance. This work was supported by grants from the Swedish Science Council, the Swedish Cancer Society, the Swedish Children's Cancer Society, STINT, the Swedish Institute, the Hjalmar Svensson Fund, and the Medical Faculty at Göteborg University. FW was partly supported by the Swedish Institute.

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Authors and Affiliations

  1. Children's Cancer Institute Australia for Medical Research, Randwick, New South Wales, Australia

    F Watt

  2. Department of Medical Chemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, Göteborg University, Gothenburg, Sweden

    F Watt, R Watanabe, W Yang, N Ågren, Y Arvidsson & K Funa

  3. Department of Medical Cell Biology, Biomedical Center, Uppsala University, Uppsala, Sweden

    N Ågren

  4. Lundberg Laboratory for Cancer Research, Sahlgrenska Academy, Göteborg University, Gothenburg, Sweden

    Y Arvidsson

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  1. F Watt
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  6. K Funa
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Correspondence to K Funa.

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Watt, F., Watanabe, R., Yang, W. et al. A novel MASH1 enhancer with N-myc and CREB-binding sites is active in neuroblastoma. Cancer Gene Ther 14, 287–296 (2007). https://doi.org/10.1038/sj.cgt.7701012

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