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Detection of novel mRNA splice variants of human ING4 tumor suppressor gene

Oncogene volume 26, pages 5247–5257 (2007)Cite this article

Abstract

Inhibitor of growth (ING)4, member of a gene family encoding potential tumor suppressors, is implicated as a repressor of angiogenesis and tumor growth and suppresses loss of contact inhibition in vitro. Here, we report that ING4 undergoes alternative splicing. Expression analysis identified novel ING4 spliced variant mRNAs encoding proteins devoid of different portions. The ING4 variants were detected in both normal and tumor tissues. The existence of ING4 variants was confirmed by several approaches, including reverse transcriptase–polymerase chain reaction, real-time PCR and in silico experiments. To investigate the functional consequences of alternative splicing the ING4 variant cDNAs were expressed in mammalian cells. Our studies indicated that (i) the ING4 variants do not differ from wild-type in their nuclear localization, interaction with p53 and association to HBO1 complex; and (ii) the ING4-ΔEx6A variant, devoid of the C-terminal portion, loses the capability to inhibit NF-κB. On the whole our data suggest that alternative splicing could modulate the activity of ING4 tumor suppressor protein.

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Acknowledgements

We thank Dr Gabriella Sozzi (Istituto Nazionale Tumori, Milan) for providing with two RNA samples, Dr Domenico Delia (Istituto Nazionale Tumori, Milan) for the GST-p53 plasmid and Dr Anthony J Capobianco (The Wistar Institute, Philadelphia, PA, USA) for CMV-p65 plasmid. We also thank the realtime PCR service at IFOM Foundation-Milan for their assistance in TaqMan experiments, and Sonia Pagliardini for technical support. This work was funded by AIRC (Italian Association for Cancer Research).

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Author notes

  1. G Raho and C Miranda: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Experimental Oncology, Operative Unit Molecular Mechanisms of Cancer Growth and Progression, Istituto Nazionale Tumori, Milan, Italy

    G Raho, C Miranda & A Greco

  2. Department of Pathology, Experimental Molecular Pathology, Istituto Nazionale Tumori, Milan, Italy

    E Tamborini

  3. Scientific Direction, Istituto Nazionale Tumori, Milan, Italy

    M A Pierotti

  4. IFOM Foundation (The Firc Institute for Molecular Oncology Foundation), Milan, Italy

    M A Pierotti

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  1. G Raho
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  2. C Miranda
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  3. E Tamborini
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  4. M A Pierotti
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  5. A Greco
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Corresponding authors

Correspondence to M A Pierotti or A Greco.

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Raho, G., Miranda, C., Tamborini, E. et al. Detection of novel mRNA splice variants of human ING4 tumor suppressor gene. Oncogene 26, 5247–5257 (2007). https://doi.org/10.1038/sj.onc.1210335

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