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Neuroblastoma-specific expression of potential therapeutics cannot be achieved using a promoter region of the NCX (TLX2) gene

Cancer Gene Therapy volume 17, pages 373–374 (2010)Cite this article

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Neuroblastoma is a pediatric solid tumor that arises from precursor cells of the sympathetic nervous system.1 Narita et al.2 published an interesting report that the promoter region of the TLX2 (NCX, Hox11L1, ENX) gene, a member of the HOX11 homeobox gene family, might be used to selectively express potential therapeutic gene sequences in neuroblastoma cells, allowing targeted expression of a suicide gene in cancer cells. The approach of conditional expression of a gene therapy has been successfully developed and tested using a variety of disease- or cell-specific promoters.3, 4 The rationale for using the TLX2 gene promoter to achieve neuroblastoma-specific expression was that expression of the endogenous TLX2 gene is primarily confined to neural crest-derived tissues.2, 5, 6, 7

The NCX1.7-luc vector is a modified pGL2-basic vector containing a 1.7-kb region of the NCX promoter upstream of the ATG start codon, which was generated through sequential deletions of genomic DNA of the NCX gene.2 This promoter region was activated in neuroblastoma cells and expression was measured as a function of luciferase expression. Notably, Narita et al. demonstrated that transfection of this construct into six neuroblastoma cell lines (SH-SY5Y, NGP, NMB, NBL-S, NLF and SMS-SAN) resulted in luciferase expression, but not in negative controls, which included fibroblasts (HFF, MRC-5 and CDC18Co), melanoma (A875), pancreatic cancer (AsPC-1) and lung carcinoma (QG56) cell lines. Our group has identified miRNAs that have anti-proliferative effects when ectopically up-regulated in neuroblastoma cell lines.8, 9 Therefore, we were interested in validating the cell specificity of NCX1.7-luc expression for use in microRNA-mediated therapeutics. The NCX1.7-luc vector was transfected into nine neuroblastoma cell lines (Kelly, SKNDZ, NGP, IMR-32, SKNBE, SKNAS, LAN-6, SH-SY5Y and SHEP T21N (a non-MYCN amplified cell line containing a tetracycline repressible MYCN expression construct)), and in several putative negative controls, including human embryonic kidney (HEK293) cells, cervical cancer (HeLa) cells, breast cancer (MDA-MB-231 and Hs578T) cells and human astrocytoma glioblastoma (U373MG) cells.

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Acknowledgements

The NCX1.7-luc vector was obtained from Dr Masahiko Hatano, Department of Biomedical Science, Chiba University, Japan. HeLa cells were kindly donated by Dr Rowan Higgs, MCT, RCSI. Hs578T cells were obtained from Ms Leanne Moore, The Ocular Genetics Unit, Trinity College Dublin, and U373MG cells were donated by Ms Elif Dagdan, The Ocular Genetics Unit, Trinity College Dublin. This work was supported by grants from Science Foundation Ireland, Children's Medical and Research Foundation, and the NIH (5R01CA127496).

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  1. Department of Cancer Genetics, Royal College of Surgeons in Ireland, Dublin, Ireland

    A Tivnan & R L Stallings

  2. Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland

    A Tivnan & R L Stallings

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Correspondence to R L Stallings.

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Tivnan, A., Stallings, R. Neuroblastoma-specific expression of potential therapeutics cannot be achieved using a promoter region of the NCX (TLX2) gene. Cancer Gene Ther 17, 373–374 (2010). https://doi.org/10.1038/cgt.2009.90

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