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Bryostatin-1 causes radiosensitization of BMG-1 malignant glioma cells through differential activation of protein kinase-Cδ not evident in the non-malignant AA8 fibroblasts

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Abstract

Bryostatin-1 (bryo-1), a non-phorbol ester, is known to sensitize mammalian cells against certain chemotherapeutic drugs. We assessed its ability to modify radiation response of mammalian cells using Chinese hamster fibroblasts AA8 cells and human malignant glioma BMG-1 cells. In the malignant glioma BMG-1 cell line, bryo-1 pre-treatment significantly enhanced radiation-induced growth inhibition and cytogenetic damage, and further reduced the clonogenic cell survival as compared to cells irradiated at the clinically relevant dose of 2 Gy. PKCδ expression increased significantly when bryo-1 pre-treated BMG-1 glioma cells were irradiated at 2 Gy and induced prolonged ERK-1/2 activation associated with p21 overexpression. Silencing PKCδ resulted in inhibition of bryo-1-induced radiosensitization. In contrast, bryo-1 failed to alter radiosensitivity (cell survival; growth inhibition; cytogenetic damage) or activate ERK1/2 pathway in the AA8 fibroblasts despite PKCδ phosphorylation at its regulatory (Y155) domain, indicating alternate mechanisms in these non-malignant cells as compared to the glioma cells. This study suggests that bryo-1 may effectively enhance the radiosensitivity of malignant cells and warrants further in-depth investigations to evaluate its radiosensitizing potential in various cell types.

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Acknowledgments

This study was conducted as part of the low-dose radiobiology investigations under DRDO Grant INM-311.1.5. RSD and SH are thankful to the Indian Government agencies ICMR and DRDO, respectively, for receiving research scholarships.

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  1. Natural Radiation Response Mechanisms Group, Division of Radiation Biosciences, Institute of Nuclear Medicine & Allied Sciences, Brig. S K Mazumdar Road, Timarpur, Delhi, 110054, India

    Raghubendra Singh Dagur, Shashank Hambarde & Sudhir Chandna

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  1. Raghubendra Singh Dagur
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  2. Shashank Hambarde
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  3. Sudhir Chandna
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Correspondence to Sudhir Chandna.

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11010_2014_2291_MOESM1_ESM.tif

Fig.S1 Radiosensitivity of BMG-1 and AA8 cells is unaffected by Bryo-1 pre-treatment at low-dose radiation. Cell proliferation kinetics of (A) BMG-1 glioma cells and (B) AA8 fibroblasts shows no effect of bryo-1 pre-treatment following irradiation at doses 0.1 Gy, 0.4 Gy, and 1.0 Gy. Results are mean  ± SD of three independent experiments. (TIFF 492 kb)

11010_2014_2291_MOESM2_ESM.tif

Fig.S2 Bryo-1 treatment prior to irradiation results in sustained activation (phosphorylation) of ERK1/2 in BMG-1 cells. Western blot shows sustained increase in the phosphorylated form of ERK1/2 in bryo-1 pre-treated BMG-1 cells at 24 h post-irradiation. (TIFF 143 kb)

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Dagur, R.S., Hambarde, S. & Chandna, S. Bryostatin-1 causes radiosensitization of BMG-1 malignant glioma cells through differential activation of protein kinase-Cδ not evident in the non-malignant AA8 fibroblasts. Mol Cell Biochem 401, 49–59 (2015). https://doi.org/10.1007/s11010-014-2291-0

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