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The synergistic effect of BCR signaling inhibitors combined with an HDAC inhibitor on cell death in a mantle cell lymphoma cell line

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Abstract

Mantle cell lymphoma (MCL) is a B cell malignancy characterized by aberrant expression of cyclin D1 due to a t(11;14) translocation. MCL is refractory to conventional chemotherapy, and treatment remains challenging. We investigated the efficacy of the histone deacetylase (HDAC) inhibitor vorinostat combined with one of several B-cell receptor (BCR) signaling inhibitors on MCL cell death and the underlying mechanisms, using MCL cell lines. The Bruton’s tyrosine kinase inhibitor PCI-32765 and the spleen tyrosine kinase inhibitor R406 showed synergistic effects with vorinostat on growth inhibition. Treatment with PCI-32765 or R406 alone induced 27.3 ± 2.1 or 25.1 ± 3.2 % apoptosis. When combined with vorinostat, these apoptotic fractions significantly increased to 50.8 ± 4.9 and 63.1 ± 5.0 %, respectively. Activation of caspase-3 and poly-(ADP-ribose) polymerase cleavage were markedly increased. We performed gene expression profiling following treatment with the combination of vorinostat and individual BCR signaling inhibitors using a microarray, and differentially expressed genes were identified. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the nuclear factor (NF)-κB signaling pathway was significantly enriched following treatment with the combination of vorinostat and R406. Protein expression analysis confirmed the down-regulation of NF-κB1/p105 and cyclin D1, suggesting inhibition of the NF-κB pathway. Taken together, the combination of an HDAC inhibitor and a BCR signaling inhibitor may be a novel therapeutic strategy for MCL.

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Abbreviations

BCR:

B-cell receptor

Btk:

Bruton’s tyrosine kinase

CI:

Combination index

DEGs:

Differentially expressed genes

DMSO:

Dimethyl sulfoxide

HDAC:

Histone deacetylase

KEGG:

Kyoto Encyclopedia of Genes and Genomes

MCL:

Mantle cell lymphoma

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NF-κB:

Nuclear factor-κB

PARP:

Poly-(ADP-ribose) polymerase

PI:

Propidium iodide

PI3K:

Phosphatidylinositol 3-kinase

Syk:

Spleen tyrosine kinase

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Acknowledgments

This work was supported by the National Cancer Center Research and Development Fund (26-A-4) and the Health and Labour Sciences Research Grant from the Ministry of Health, Labour, and Welfare of Japan (H25-AIDS-I-002).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Clinical Research Center, National Hospital Organization Nagoya Medical Center, 4-1-1, Sannomaru, Naka-ku, Nagoya, Japan

    Kazumi Hagiwara, Shinji Kunishima, Hiroatsu Iida, Yasuhiko Miyata & Hirokazu Nagai

  2. National Hospital Organization Nagoya Medical Center, 4-1-1, Sannomaru, Naka-ku, Nagoya, Japan

    Tomoki Naoe

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  1. Kazumi Hagiwara
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  2. Shinji Kunishima
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  3. Hiroatsu Iida
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  6. Hirokazu Nagai
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Correspondence to Hirokazu Nagai.

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Hagiwara, K., Kunishima, S., Iida, H. et al. The synergistic effect of BCR signaling inhibitors combined with an HDAC inhibitor on cell death in a mantle cell lymphoma cell line. Apoptosis 20, 975–985 (2015). https://doi.org/10.1007/s10495-015-1125-1

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