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Functional inhibition of BCL2 is needed to increase the susceptibility to apoptosis to SMO inhibitors in diffuse large B-cell lymphoma of germinal center subtype

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Abstract

Previously, we have demonstrated that inhibition of Hedgehog pathway induces predominantly apoptosis in diffuse large B-cell lymphoma (DLBCL) cell lines of activated B-cell (ABC) type but predominantly cell cycle arrest in those of germinal center (GC). Here, we explored the possibility of overcoming the resistance to apoptosis to SMO inhibitors in five DLBCL cells of GC type using the combination of the SMO inhibitor HhAntag (Genentech Inc) with the BH3 mimetic ABT-737 (Abbott Laboratories). As controls we have used two DLBCL of ABC type (OCI-LY10 and OCI-LY3). Combinatorial treatments were performed with increasing concentrations of the HhAntag with low doses (equal or less than the IC20) of ABT-737. MTS assays were used to detect changes in cell viability and Annexin-V and PARP1 cleavage assays were used to detect apoptosis. Combining low doses of ABT-737 with increasing concentrations of HhAntag in GC DLBCL cell lines resulted in significantly increase of apoptosis in comparison to treatments with the SMO inhibitor alone. We concluded that in GC DLBCL cell lines, in contrast to those of ABC type, functional inhibition of BCL2 family members is usually needed to overcome the resistance to apoptosis to SMO inhibitors. These findings provide a rationale to explore the use of SMO and BCL2 inhibitors as adjuvant therapy for treatment of DLBCL of GC type.

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Acknowledgments

This work was supported by funds from The Translational Grant of The Leukemia and Lymphoma Society (to RS and FV), and K08 Physician-Scientist Award 1K08 CA143151-01 (NIH) (to FV), and SPORE Lymphoma Grant 1P50CA136411-01A1 (to FV).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Unit 72, 1515 Holcombe Boulevard, Houston, TX, 77030, USA

    Kranthi Kunkalla, Yadong Liu, Changju Qu, Vasiliki Leventaki, Nitin K. Agarwal, Rajesh R. Singh & Francisco Vega

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  1. Kranthi Kunkalla
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  2. Yadong Liu
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  3. Changju Qu
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  4. Vasiliki Leventaki
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  6. Rajesh R. Singh
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  7. Francisco Vega
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Correspondence to Francisco Vega.

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Supplementary figure 1

Combining low doses of ABT-737 and increasing concentrations of HhAntag resulted in decreased cell viability in Toledo cells. Using the cell count assay Vi-Cell viability analyzer (Beckman coulter) showed that combining IC20-doses of ABT-737 with increasing concentrations of HhAntag for 48 h resulted in decreased cell viability in comparison with treatments of HhAntag alone in Toledo cells confirming the findings seen in the Annexin PI assay. The minimally lethal effect of the concentration of ABT-737 used (10 nM) for the combinatorial treatment is evident by the little decrease in the cell viability after treatment with ABT-737 alone for 48 h (left panels). (PPT 35 kb)

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Kunkalla, K., Liu, Y., Qu, C. et al. Functional inhibition of BCL2 is needed to increase the susceptibility to apoptosis to SMO inhibitors in diffuse large B-cell lymphoma of germinal center subtype. Ann Hematol 92, 777–787 (2013). https://doi.org/10.1007/s00277-013-1684-6

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  • DOI: https://doi.org/10.1007/s00277-013-1684-6

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