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Genotype-dependent cooperation of ionizing radiation with BRAF inhibition in BRAF V600E-mutated carcinomas

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Summary

Background A substantial proportion of solid tumors carry the BRAF V600E mutation, which causes activation of the MEK/MAPK pathway and is a poor prognostic indicator. Patients with locally advanced human cancers are often treated with external beam radiation therapy. Given the association of Raf overactivation with radioresistance, we hypothesized that, in BRAF V600E-mutated carcinomas, there would be combinatorial activity between radiation and PLX4720, a specific BRAF V600E-inhibitor. Methods Two BRAF V600E-mutated cancer cell lines and one BRAF-V600E wildtype (WT) cancer cell line were obtained. We performed cell viability assays and clonogenic assays using combinations of radiation and PLX4720. We assessed MEK and MAPK phosphorylation at different PLX4720 concentrations with western blotting, and cell cycle progression was evaluated by flow cytometry. Results Our results show combinatorial, additive activity between radiation and PLX4720 in BRAF V600E-mutated cell lines, but not in the BRAF WT line. In BRAF V600E-mutated cells, there was a PLX4720 concentration-dependent decrease in MEK and MAPK phosphorylation. In cells with BRAF V600E mutations, PLX4720 caused cell cycle arrest at G1, and, when combined with radiation, caused a combined G1 and G2 cell cycle arrest; this pattern of cell cycle effects was not seen in the BRAF WT cell line. Conclusions These data suggest additive, combinatorial activity between radiation and PLX4720 in cancers carrying BRAF V600E mutations. Our data has potential for translation into the multimodality treatment of BRAF V600E-mutated cancers.

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Acknowledgments

We thank Plexxikon, Inc. for providing PLX4720, and Dr. Brian L. West for this thoughtful advice and assistance during this study.

Author disclosures and conflict of interest

None. The authors report no conflict of interest.

Ethical standards

The submitted manuscript complies with the current laws of the country, the United States of America, in which the experiments and analysis were performed.

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Authors and Affiliations

  1. Department of Radiation Oncology, University of California, San Francisco, 1600 Divisadero Street, Suite H-1031, San Francisco, CA, 94143-1708, USA

    Tina Dasgupta, Daphne A. Haas-Kogan, Xiaodong Yang, Aleksandra Olow, Daniel X. Yang, Ashley Gragg & Sue S. Yom

  2. Department of Otolaryngology - Head and Neck Surgery, University of California, San Francisco, 2380 Sutter Street, 2nd Floor, Box 1703, San Francisco, CA, 94143-1703, USA

    Lisa A. Orloff & Sue S. Yom

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  1. Tina Dasgupta
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  2. Daphne A. Haas-Kogan
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  3. Xiaodong Yang
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Correspondence to Sue S. Yom.

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Dasgupta, T., Haas-Kogan, D.A., Yang, X. et al. Genotype-dependent cooperation of ionizing radiation with BRAF inhibition in BRAF V600E-mutated carcinomas. Invest New Drugs 31, 1136–1141 (2013). https://doi.org/10.1007/s10637-013-9928-9

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  • DOI: https://doi.org/10.1007/s10637-013-9928-9

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