Effects of carbon ion irradiation and X-ray irradiation on the ubiquitylated protein accumulation

Isozaki,Tetsuro; Fujita,Mayumi; Yamada,Shigeru; Imadome,Kaori; Shoji,Yoshimi; Yasuda,Takeshi; Nakayama,Fumiaki; Imai,Takashi; Matsubara,Hisahiro

doi:10.3892/ijo.2016.3504

Effects of carbon ion irradiation and X-ray irradiation on the ubiquitylated protein accumulation

Authors:
- Tetsuro Isozaki
- Mayumi Fujita
- Shigeru Yamada
- Kaori Imadome
- Yoshimi Shoji
- Takeshi Yasuda
- Fumiaki Nakayama
- Takashi Imai
- Hisahiro Matsubara
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Affiliations: Department of Frontier Surgery, Chiba University Graduate School of Medicine, Chiba, Japan, Advanced Radiation Biology Research Program, National Institute of Radiological Sciences, Chiba, Japan, Research Center Hospital, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba, Japan, Radiation Emergency Medicine Research Program, Research Center for Radiation Emergency Medicine, National Institute of Radiological Sciences, Chiba, Japan
Published online on: May 5, 2016 https://doi.org/10.3892/ijo.2016.3504
Pages: 144-152
Copyright: © Isozaki et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

C-ion radiotherapy is associated with improved local control and survival in several types of tumors. Although C-ion irradiation is widely reported to effectively induce DNA damage in tumor cells, the effects of irradiation on proteins, such as protein stability or degradation in response to radiation stress, remain unknown. We aimed to compare the effects of C-ion and X-ray irradiation focusing on the cellular accumulation of ubiquitylated proteins. Cells from two human colorectal cancer cell lines, SW620 and SW480, were subjected to C-ion or X-ray irradiation and determination of ubiquitylated protein levels. High levels of ubiquitylated protein accumulation were observed in the C-ion-irradiated SW620 with a peak at 3 Gy; the accumulation was significantly lower in the X-ray-irradiated SW620 at all doses. Enhanced levels of ubiquitylated proteins were also detected in C-ion or X-ray-irradiated SW480, however, those levels were significantly lower than the peak detected in the C-ion-irradiated SW620. The levels of irradiation-induced ubiquitylated proteins decreased in a time-dependent manner, suggesting that the proteins were eliminated after irradiation. The treatment of C-ion-irradiated SW620 with a proteasome inhibitor (epoxomicin) enhanced the cell killing activity. The accumulated ubiquitylated proteins were co-localized with γ-H2AX, and with TP53BP1, in C-ion-irradiated SW620, indicating C-ion-induced ubiquitylated proteins may have some functions in the DNA repair system. Overall, we showed C-ion irradiation strongly induces the accumulation of ubiquitylated proteins in SW620. These characteristics may play a role in improving the therapeutic ratio of C-ion beams; blocking the clearance of ubiquitylated proteins may enhance sensitivity to C-ion radiation.

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