Hyperthermia-induced radiosensitization in CHO wild-type, NHEJ repair mutant and HR repair mutant following proton and carbon-ion exposure

Maeda,Junko; Fujii,Yoshihiro; Fujisawa,Hiroshi; Hirakawa,Hirokazu; Cartwright,Ian  M.; Uesaka,Mitsuru; Kitamura,Hisashi; Fujimori,Akira; Kato,Takamitsu  A.

doi:10.3892/ol.2015.3732

Hyperthermia-induced radiosensitization in CHO wild-type, NHEJ repair mutant and HR repair mutant following proton and carbon-ion exposure

Authors:
- Junko Maeda
- Yoshihiro Fujii
- Hiroshi Fujisawa
- Hirokazu Hirakawa
- Ian M. Cartwright
- Mitsuru Uesaka
- Hisashi Kitamura
- Akira Fujimori
- Takamitsu A. Kato
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Affiliations: Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO 80523, USA, Department of Radiological Sciences, Ibaraki Prefectural University of Health Sciences, Inashiki, Ibaraki 300‑0394, Japan, School of Engineering, The University of Tokyo, Tokyo 113‑8656, Japan, Research Center for Charged Particle Therapy, International Open Laboratory, National Institute of Radiological Sciences, Chiba 263‑8555, Japan, Research Development and Support Center, National Institute of Radiological Sciences, Chiba 263‑8555, Japan
Published online on: September 21, 2015 https://doi.org/10.3892/ol.2015.3732
Pages: 2828-2834
Copyright: © Maeda et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The DNA repair mechanisms involved in hyperthermia-induced radiosensitization with proton and carbon ion radiation exposure were investigated in the present study. In a previous study, Chinese hamster ovary (CHO) cells were exposed to low linear energy transfer (LET) photon radiation. These cells can be sensitized by hyperthermia as a result of inhibition of homologous recombination (HR) repair. The present study used wild‑type, non‑homologous end joining (NHEJ) and HR repair‑deficient CHO cells to define the contributions of each repair pathway to cellular lethality following hyperthermia‑induced hadron radiation sensitization. The cells were exposed to ionizing radiation, followed by hyperthermia treatment (42.5˚C for 1 h). Hyperthermia‑induced radiosensitization was determined by the colony formation assay and thermal enhancement ratio. HR repair‑deficient cells exhibited no hyper‑sensitization to X‑rays, protons, or low and high LET carbon ions when combined with hyperthermia. Wild‑type and NHEJ repair‑deficient cells exhibited significant hyperthermia‑induced sensitization to low LET photon and hadron radiation. Hyperthermia‑induced sensitization to high LET carbon‑ion radiation was less than at low LET radiation. Relative biological effectiveness (RBE) between radiation alone and radiation combined with hyperthermia cell groups was not significantly different in any of the cell lines, with the exception of wild‑type cells exposed to high LET radiation, which exhibited a lower RBE in the combined group. The present study investigated additional cell lines to confirm the lower RBE observed in DNA repair‑deficient cell lines. These findings suggested that hyperthermia‑induced hyper‑sensitization to hadron radiation is also dependent on inhibition of HR repair, as was observed with photon radiation in a previous study.

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1	Dewey WC: Arrhenius relationships from the molecule and cell to the clinic. Int J Hyperther. 25:3–20. 2009. View Article : Google Scholar
2	Gerweck LE, Gillette EL and Dewey WC: Effect of heat and radiation on synchronous Chinese hamster cells: Killing and repair. Radiat Res. 64:611–623. 1975. View Article : Google Scholar : PubMed/NCBI
3	Holahan EV, Highfield DP, Holahan PK and Dewey WC: Hyperthermic killing and hyperthermic radiosensitization in Chinese hamster ovary cells: Effects of pH and thermal tolerance. Radiat Res. 97:108–131. 1984. View Article : Google Scholar : PubMed/NCBI
4	Dewey WC and Miller HH: Effect of temperature on x-ray induced cell lethality and chromosomal aberrations. Int J Radiat Biol Relat Stud Phys Chem Med. 18:91–93. 1970. View Article : Google Scholar : PubMed/NCBI
5	Lee YJ and Dewey WC: Induction of heat shock proteins in Chinese hamster ovary cells and development of thermotolerance by intermediate concentrations of puromycin. J Cell Physiol. 132:1–11. 1987. View Article : Google Scholar : PubMed/NCBI
6	Lee YJ and Dewey WC: Effect of cycloheximide or puromycin on induction of thermotolerance by heat in Chinese hamster ovary cells: Dose fractionation at 45.5 degrees C1. Cancer Res. 47:5960–5966. 1987.PubMed/NCBI
7	Genet SC, Fujii Y, Maeda J, Kaneko M, Genet MD, Miyagawa K and Kato TA: Hyperthermia inhibits homologous recombination repair and sensitizes cells to ionizing radiation in a time- and temperature-dependent manner. J Cell Physiol. 228:1473–1481. 2013. View Article : Google Scholar : PubMed/NCBI
8	Krawczyk PM, Eppink B, Essers J, Stap J, Rodermond H, Odijk H, Zelensky A, van Bree C, Stalpers LJ, Buist MR, et al: Mild hyperthermia inhibits homologous recombination, induces BRCA2 degradation, and sensitizes cancer cells to poly (ADP-ribose) polymerase-1 inhibition. Proc Natl Acad Sci USA. 108:9851–9856. 2011. View Article : Google Scholar : PubMed/NCBI
9	Kamada T, Tsujii H, Tsuji H, Yanagi T, Mizoe JE, Miyamoto T, Kato H, Yamada S, Morita S, Yoshikawa K, et al: Working Group for the Bone and Soft Tissue Sarcomas: Efficacy and safety of carbon ion radiotherapy in bone and soft tissue sarcomas. J Clin Oncol. 20:4466–4471. 2002. View Article : Google Scholar : PubMed/NCBI
10	Tsujii H and Kamada T: A review of update clinical results of carbon ion radiotherapy. Jpn J Clin Oncol. 42:670–685. 2012. View Article : Google Scholar : PubMed/NCBI
11	Takahashi A, Ohnishi K, Wang X, Kobayashi M, Matsumoto H, Tamamoto T, Aoki H, Furusawa Y, Yukawa O and Ohnishi T: The dependence of p53 on the radiation enhancement of thermosensitivity at different let. Int J Radiat Oncol Biol Phys. 47:489–494. 2000. View Article : Google Scholar : PubMed/NCBI
12	Chang PY, Tobias CA and Blakely EA: Protein synthesis modulates the biological effectiveness of the combined action of hyperthermia and high-LET radiation. Radiat Res. 129:272–280. 1992. View Article : Google Scholar : PubMed/NCBI
13	Kinashi Y, Masunaga SI, Suzuki M, Ono K and Ohnishi T: Hyperthermia enhances thermal-neutron-induced cell death of human glioblastoma cell lines at low concentrations of 10B. Int J Radiat Oncol Biol Phys. 40:1185–1192. 1998. View Article : Google Scholar : PubMed/NCBI
14	Takahashi A, Ohnishi K, Ota I, Asakawa I, Tamamoto T, Furusawa Y, Matsumoto H and Ohnishi T: p53-dependent thermal enhancement of cellular sensitivity in human squamous cell carcinomas in relation to LET. Int J Radiat Biol. 77:1043–1051. 2001. View Article : Google Scholar : PubMed/NCBI
15	Stackhouse MA and Bedford JS: An ionizing radiation-sensitive mutant of CHO cells: Irs-20. I. Isolation and initial characterization. Radiat Res. 136:241–249. 1993. View Article : Google Scholar : PubMed/NCBI
16	Jeggo PA and Kemp LM: X-ray-sensitive mutants of Chinese hamster ovary cell line. Isolation and cross-sensitivity to other DNA-damaging agents. Mutat Res. 112:313–327. 1983.PubMed/NCBI
17	Whitmore GF, Varghese AJ and Gulyas S: Cell cycle responses of two X-ray sensitive mutants defective in DNA repair. Int J Radiat Biol. 56:657–665. 1989. View Article : Google Scholar : PubMed/NCBI
18	Fuller LF and Painter RB: A Chinese hamster ovary cell line hypersensitive to ionizing radiation and deficient in repair replication. Mutat Res. 193:109–121. 1988.PubMed/NCBI
19	Hinz JM, Tebbs RS, Wilson PF, Nham PB, Salazar EP, Nagasawa H, Urbin SS, Bedford JS and Thompson LH: Repression of mutagenesis by Rad51D-mediated homologous recombination. Nucleic Acids Res. 34:1358–1368. 2006. View Article : Google Scholar : PubMed/NCBI
20	Kato TA, Tsuda A, Uesaka M, Fujimori A, Kamada T, Tsujii H and Okayasu R: In vitro characterization of cells derived from chordoma cell line U-CH1 following treatment with X-rays, heavy ions and chemotherapeutic drugs. Radiat Oncol. 6:1162011. View Article : Google Scholar : PubMed/NCBI
21	Maeda J, Roybal EJ, Brents CA, Uesaka M, Aizawa Y and Kato TA: Natural and glucosyl flavonoids inhibit poly(ADP-ribose) polymerase activity and induce synthetic lethality in BRCA mutant cells. Oncol Rep. 31:551–556. 2014.PubMed/NCBI
22	Zeng ZC, Jiang GL, Wang GM, Tang ZY, Curran WJ and Iliakis G: DNA-PKcs subunits in radiosensitization by hyperthermia on hepatocellular carcinoma hepG2 cell line. World J Gastroenterol. 8:797–803. 2002.PubMed/NCBI
23	Ihara M, Takeshita S, Okaichi K, Okumura Y and Ohnishi T: Heat exposure enhances radiosensitivity by depressing DNA-PK kinase activity during double strand break repair. Int J Hyperthermia. 30:102–109. 2014. View Article : Google Scholar : PubMed/NCBI
24	Tomita M: Involvement of DNA-PK and ATM in radiation- and heat-induced DNA damage recognition and apoptotic cell death. J Radiat Res (Tokyo). 51:493–501. 2010. View Article : Google Scholar
25	Umeda N, Matsumoto Y, Yin HL, Tomita M, Enomoto A, Morita A, Mizukoshi T, Sakai K, Hosoi Y and Suzuki N: Difference in the heat sensitivity of DNA-dependent protein kinase activity among mouse, hamster and human cells. Int J Radiat Biol. 79:671–680. 2003. View Article : Google Scholar : PubMed/NCBI
26	Woudstra EC, Konings AW, Jeggo PA and Kampinga HH: Role of DNA-PK subunits in radiosensitization by hyperthermia. Radiat Res. 152:214–218. 1999. View Article : Google Scholar : PubMed/NCBI
27	Weyrather WK, Ritter S, Scholz M and Kraft G: RBE for carbon track-segment irradiation in cell lines of differing repair capacity. Int J Radiat Biol. 75:1357–1364. 1999. View Article : Google Scholar : PubMed/NCBI
28	Olsson G, Czene S, Jenssen D and Harms-Ringdahl M: Induction of homologous recombination in the hprt gene of V79 Chinese hamster cells in response to low- and high-LET irradiation. Cytogenet Genome Res. 104:227–231. 2004. View Article : Google Scholar : PubMed/NCBI
29	Zafar F, Seidler SB, Kronenberg A, Schild D and Wiese C: Homologous recombination contributes to the repair of DNA double-strand breaks induced by high-energy iron ions. Radiat Res. 173:27–39. 2010. View Article : Google Scholar : PubMed/NCBI
30	Genet SC, Maeda J, Fujisawa H, Yurkon CR, Fujii Y, Romero AM, Genik PC, Fujimori A, Kitamura H and Kato TA: Comparison of cellular lethality in DNA repair-proficient or -deficient cell lines resulting from exposure to 70 MeV/n protons or 290 MeV/n carbon ions. Oncol Rep. 28:1591–1596. 2012.PubMed/NCBI
31	Mitchel RE, Smith BP, Wheatly N, Chan A, Child S and Paterson MC: Sensitivity of hyperthermia-treated human cells to killing by ultraviolet or gamma radiation. Radiat Res. 104:234–241. 1985. View Article : Google Scholar : PubMed/NCBI
32	Mitchel RE, Chan A, Smith BP, Child SD and Paterson MC: The effects of hyperthermia and ionizing radiation in normal and ataxia telangiectasia human fibroblast lines. Radiat Res. 99:627–635. 1984. View Article : Google Scholar : PubMed/NCBI