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Targeting NF-κB in infantile hemangioma-derived stem cells reduces VEGF-A expression

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Abstract

Background

Infantile hemangioma (IH) is a most common tumor of infancy. Using infantile hemangioma-derived stem cells (HemSCs), we recently demonstrated that corticosteroids suppress the expression of VEGF-A, monocyte chemoattractant protein-1 (MCP-1), urokinase plasminogen activator receptor (uPAR), and interleukin-6 (IL-6); each of these are known targets of the transcription factor nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB). In the present study, we examined the expression of these NF-κB target genes in IH tissue specimens and the effect of NF-κB regulation on the expression of pro-angiogenic cytokines, and in particular VEGF-A, in HemSCs.

Materials and methods

RNA extracted from IH tissue and hemangioma-derived stem cells (HemSCs) was used to analyze NF-κB target gene expression by reverse transcription–quantitative PCR (RT-qPCR). The effects of NF-κB blockade were examined in HemSCs. Immunostaining, immunoblotting and ELISA were used to assess protein expression.

Results

MCP-1, uPAR, and IL-6 were found to be differentially expressed in proliferating versus involuting IH. Corticosteroids suppressed NF-κB activity of HemSCs. Velcade (Bortezomib), a proteosome inhibitor that can indirectly inhibit NF-κB, impaired HemSCs viability and expression of pro-angiogenic factors. Furthermore, specific inhibition of NF-κB resulted in suppression of VEGF-A.

Conclusions

We demonstrate expression of NF-κB target genes in proliferating IH. In addition, we show that the expression of several pro-angiogenic factors in HemSCs, and in particular VEGF–A, is regulated by NF-B activity.

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Acknowledgments

We thank Debajit K. Biswas, Department of Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, for invaluable advice and for providing NBD peptide for initial experiments. Supported by a NIH R01 HL096384 (JB), the Talpiot Medical Leadership Program, Sheba Medical Center, Israel (S.G.), Harvard Skin Diseases Pilot Study Grant (S.G.), and the John Butler Mulliken Foundation.

Conflict of Interest

The authors have declared that no conflict of interest exists.

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Authors and Affiliations

  1. Vascular Biology Program and Department of Surgery, Children’s Hospital Boston and Harvard Medical School, Boston, MA, 02115, USA

    Shoshana Greenberger, Irit Adini, Elisa Boscolo & Joyce Bischoff

  2. Department of Plastic Surgery, Children’s Hospital Boston and Harvard Medical School, Boston, MA, 02115, USA

    John B. Mulliken

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  1. Shoshana Greenberger
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  2. Irit Adini
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  3. Elisa Boscolo
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  4. John B. Mulliken
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Correspondence to Joyce Bischoff.

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Greenberger, S., Adini, I., Boscolo, E. et al. Targeting NF-κB in infantile hemangioma-derived stem cells reduces VEGF-A expression. Angiogenesis 13, 327–335 (2010). https://doi.org/10.1007/s10456-010-9189-6

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  • DOI: https://doi.org/10.1007/s10456-010-9189-6

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