Inhibition of DNA-PK activity and radio-sensitization induced by single strand DNA

• Project/Area Number: 15390357
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Radiation science
• Research Institution: The University of Tokyo
• Principal Investigator: HOSOI Yoshio The University of Tokyo, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (50238747)
• Project Period (FY): 2003 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥13,700,000 (Direct Cost: ¥13,700,000); Fiscal Year 2005: ¥4,500,000 (Direct Cost: ¥4,500,000); Fiscal Year 2004: ¥4,500,000 (Direct Cost: ¥4,500,000); Fiscal Year 2003: ¥4,700,000 (Direct Cost: ¥4,700,000)
• Keywords: ionizing radiation / DNA-PK / single strand DNA / DNA double strand breaks / suramin / EGF受容体 / 2重鎖切断 / DNA修復 / 癌

Research Abstract

Phosphorothioate oligonucleotides and suramin bind to heparin binding proteins including DNA polymerases, and inhibit their functions. In the present study, we report inhibition of DNA-dependent protein kinase (DNA-PK) activity by phosphorothioate oligonucleotides and suramin. Inhibitory effect of phosphorothioate oligonucleotides on DNA-PK activity was increased with length and reached a plateau at 36-mer. The base composition of phosphorothioate oligonucleotides did not affect the inhibitory effect. The inhibitory effect by phosphorothioate oligodeoxycytidine 36-mer can be about 200-fold greater than that by the phosphodiester oligodeoxycytidine 36-mer. The inhibitory effect was also observed with purified DNA-PK, which suggests direct interaction between DNA-PK and phosphorothioate oligonucleotides. DNA-PK will have different binding positions for double-stranded DNA and phosphorothioate oligodeoxycytidine 36-mer because they were not competitive in DNA-PK activation. Suramin sensit ized cells to X-radiation in a dose-dependent fashion and longer exposure to suramin before X-irradiation resulted in more efficient sensitization. The dose-modifying factors calculated from the survival curves were 1.18 in LM217 cells and 1.37 in MDA-MB-468 cells. Suramin did not sensitized scid cells that had no DNA-dependent protein kinase activity. Suramin inhibited DNA-dependent protein kinase activity in vitro and in vivo. The concentration of suramin resulting in 50% inhibition in vitro was 1.7 μM in LM217 cells and 2.4 μM in MDA-MB-468 cells. Exposure of LM217 and MDA-MB-468 cells to suramin did not affect the level of Ku70 or Ku80, but it increased the level of DNA-PKcs. Suramin did not sensitize LM217 or MDA-MB-468 cells to UV radiation. Exposure to suramin inhibited the repair of DNA double-strand breaks caused by 50 Gy irradiation. Suramin's effects were not caused by accumulation of cells in a specific phase of the cell cycle. These results suggest that suramin sensitizes cells to ionizing radiation by inhibiting DNA-dependent protein kinase activity.

Research Products

• [Journal Article] Heterogeneous expression of DNA-dependent protein kinase in esophageal cancer and normal epithelium. (2006)
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  • Journal Title: International Journal of Molecular Medicine (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Src tyrosine kinase inhibitor PP2 suppresses ERK1/2 activation and epiderinal growth factor receptor transactivation by X-irradiation. (2006)
  • Author(s): Li Z, et al.
  • Journal Title: Biochemical and Biophysical Research Communications 342 Pages: 363-368
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Preservation of ovarian function by ovarian shielding when undergoing total body irradiation for hematopoietic stem cell transplantation : a report of two successful cases. (2006)
  • Author(s): Nakagawa K, et al.
  • Journal Title: Bone Marrow Transplantation 37 Pages: 583-587
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The association of DNA-dependent protein kinase activity with chromosomal instability and risk of cancer. (2006)
  • Author(s): Someya M, et al.
  • Journal Title: Carcinogenesis 27 Pages: 117-122
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Sodium orthovanadate suppresses DNA damage-induced caspase activation and apoptosis by inactivating p53. (2006)
  • Author(s): Morita A, et al.
  • Journal Title: Cell Death and Differentiation 13 Pages: 499-511
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Src tyrosine kinase inhibitor PP2 suppresses ERK1/2 activation and epidermal growth factor receptor transactivation by X-irradiation. (2006)
  • Author(s): Li Z, et al.
  • Journal Title: Biochemical and Biophysical Research Communications 342 Pages: 363-368
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Radiation therapy for portal venous invasion by hepatocellular carcinoma. (2005)
  • Author(s): Nakagawa K, et al.
  • Journal Title: World Journal of Gastroenterology 11 Pages: 7237-7241
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Effect of nitric oxide on γ-ray-induced micronucleus frequency in RAW264.7 cells. (2005)
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  • Journal Title: Radiation Research 164 Pages: 723-732
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Prognostic significance of Ku7O protein expression in patients with advanced colorectal cancer. (2005)
  • Author(s): Komuro Y, et al.
  • Journal Title: Hepato-Gasteroenterology 52 Pages: 995-998
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] アポトーシスシグナリングと放射線防護 (2005)
  • Author(s): 森田明典他
  • Journal Title: 放射線生物研究 40 Pages: 360-384
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Radiation thrapy for portal venous invasion by hepatocellular carcinoma. (2005)
  • Author(s): Nakagawa K, et al.
  • Journal Title: World Journal of Gastroenterology 11 Pages: 7237-7241
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Prognostic significance of Ku70 protein expression in patients with advanced colorectal cancer. (2005)
  • Author(s): Komuro Y, et al.
  • Journal Title: Hepato-Gasteroenterology 52 Pages: 995-998
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Apoptosis signaling and radiation protection. (2005)
  • Author(s): Morita A, et al.
  • Journal Title: Radiation Biology Research Communications 40 Pages: 360-384
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Low-dose exposures and bio-defense system. (2005)
  • Author(s): Hosoi Y.
  • Journal Title: Institute for Environmental Sciences, Japan
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Potentiality of DNA-dependent protein kinase to phosphorylate Ser46 of human p53. (2004)
  • Author(s): komiyama S, et al.
  • Journal Title: Biochemical and Biophysical Research Communications 323 Pages: 816-822
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Radiosensitization by hyperthermia in the chicken B-lymphocyte cell line DT4O and its derivatives lacking nonhomologous end joining and/or homologous recombination pathways of DNA double-strand break repair. (2004)
  • Author(s): Yin HL, et al.
  • Journal Title: Radiation Research 162 Pages: 433-441
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Spontaneous mutations in digestive tract of old mice show tissue-specific patterns of genomic instability (2004)
  • Author(s): Ono T, et al.
  • Journal Title: Cancer Research 64 Pages: 6919-6923
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Up-regulation of DNA-dependent protein kinase activity and Spl in colorectal cancer. (2004)
  • Author(s): Hosoi Y, et al.
  • Journal Title: International Journal of Oncology 25 Pages: 461-468
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Suramin sensitizes cells to ionizing radiation by inactivating DNA-dependent protein kinase. (2004)
  • Author(s): Hosoi Y, et al.
  • Journal Title: Radiation Research 162 Pages: 308-314
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 遺伝子レベルでの時間と空間 (2004)
  • Author(s): 細井義夫
  • Journal Title: 癌の臨床 50 Pages: 1055-1060
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 化学放射線療法の基礎 (2004)
  • Author(s): 細井義夫
  • Journal Title: 日本婦人科腫瘍学会誌 22 Pages: 334-342
  • NAID: 10026151968
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Potentiality of DNA-dependent protein kinase to phosphorylate Ser46 of human p53. (2004)
  • Author(s): Komiyama S, et al.
  • Journal Title: Biochemical and Biophysical Research Communications 323 Pages: 816-822
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Radiosensitization by hyperthermia in the chicken B-lymphocyte cell line DT40 and its derivatives lacking nonhomologous end joining and/or homologous recombination pathways of DNA double-strand break repair. (2004)
  • Author(s): Yin HL, et al.
  • Journal Title: Radiation Research 162 Pages: 433-441
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Spontaneous mutations in Digestive tract of old mice show tissue-specific patterns of genomic instability (2004)
  • Author(s): Ono T, et al.
  • Journal Title: Cancer Research 64 Pages: 6919-6923
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Up-regulation of DNA-dependent protein kinase activity and Sp1 in colorectal cancer. (2004)
  • Author(s): Hosoi Y, et al.
  • Journal Title: International Journal of Oncology 25 Pages: 461-468
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Radiation-induced signal transduction pathways. (2004)
  • Author(s): Hosoi Y.
  • Journal Title: Japanese Journal of Cancer Clinics 50 Pages: 1055-1060
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Basics of combined chemo-radiotherapy. (2004)
  • Author(s): Hosoi Y.
  • Journal Title: The Journal of the Japan Society of Gynecologic Oncology 22 Pages: 334-342
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Suppression of anchorage-independent growth by expression of the ataxia-telangiectasia group D complementing gene, ATDC. (2004)
  • Author(s): Hosoi, Y. et al.
  • Journal Title: Molecular Carciogenesis (In press)
• [Journal Article] Up-regulation of DNA-dependent protein kinase activity and Sp1 in colorectal cancer. (2004)
  • Author(s): Hosoi, Y. et al.
  • Journal Title: International Journal of Oncology 25 Pages: 461-468
• [Journal Article] Spontaneous mutations in Digestive tract of old mice show tissue-specific patterns of genomic instability. (2004)
  • Author(s): Ono, T., Ikehata, H., Pithani, V.P., Uehara, Y., Chan, Y., Kinouchi, Y., Shimosegawa, Y., Hosoi, Y.
  • Journal Title: Cancer Research 64 Pages: 6919-6923
• [Journal Article] Radiosensitization by hyperthermia in the chicken B-lymphocyte cell line DT40 and its derivatives lacking nonhomologous end joining and/or homologous recombination pathways of DNA bouble-strand break repair. (2004)
  • Author(s): Yin, H.L., Suzuki, Y., Matsumoto, Y., Tomita, M., Furusawa, Y., Enomoto, A., Motrita, A., Aoki, M., Yatagai, F., Suzuki, T., Hosoi, Y.et al.
  • Journal Title: Radiation Research 162 Pages: 433-441
• [Journal Article] Potentiality of DNA-dependent protein kinase to phosphorylate Ser46 of human p53. (2004)
  • Author(s): Komiyama, S., Taniguchi, S., Matsumoto, Y., Tsunoda, E., Ohto, T., Suzuki, Y., Yin, H.-L., Tomita, M., Enomoto, A., Motita, A., Suzuki, T., Ohtomo, K., Hosoi, Y.et al.
  • Journal Title: Biochemical and Biophysical Research Communications 323 Pages: 816-822
• [Journal Article] Suramin sensitizes cells to ionizing radiation by inactiveating DNA-dependent protein kinase. (2004)
  • Author(s): Hosoi, Y. et al.
  • Journal Title: Radiation Research 162 Pages: 308-314
• [Book] Low-Dose Radiation Exposures and Bio-Defense System (2005)
  • Author(s): Hosoi Y, et al.
  • Total Pages: 117
  • Publisher: Institute for Environmental Sciences, Japan
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Hosoi, Y.: "Suppression of anchorage-independent growth by expression of the ataxia-telangiectasia group D complementing gene, ATDC."Molecular Carcinogenesis. (in press).
• [Publications] Hosoi, Y.: "Suramin sensitizes cells to ionizing radiation by inactivating DNA-dependent protein kinase."Radiation Research. (in press).
• [Publications] Enomoto, A.: "Decreased c-Myc expression involved in X-ray-induced apoptotic cell death of human T-cell leukemia cell line MOLT-4."International Journal of Radiation Biology. 79. 589-600 (2003)
• [Publications] Komuro, Y.: "Prediction of tumor radiosensitivity in rectal carcinoma based on p53 and Ku70 expression."J.Exp.Clin.Cancer Res.. 22. 223-228 (2003)
• [Publications] Enomoto, A.: "Caspase-mediated cleavage of JNK during stress-induced apoptosis."Biochemical and Biophysical Research Communications. 306. 837-842 (2003)
• [Publications] Umeda N.: "Difference in the heat sensitivity of DNA-dependent protein kinase activity among mouse, hamster and human cells."International Journal of Radiation Biology. 79. 671-680 (2003)