Analysis of neoplastic cell-specific fatal effects with reactive oxygen species abnormality derived from compound from plant species, piperlongumine

• Project/Area Number: 15K00537
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Kyoto University
• Principal Investigator: Tano Keizo 京都大学, 複合原子力科学研究所, 准教授 (00183468)
• Co-Investigator(Kenkyū-buntansha): 増永 慎一郎 京都大学, 複合原子力科学研究所, 教授 (80238914)
• Research Collaborator: NAGASAWA Hideko
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: DNA損傷修復 / 細胞内微細環境 / 活性酸素種

Outline of Final Research Achievements

We assessed genotoxic potentials of Piperlongumine (PL) and Tirapazamine (TPZ) by utilizing DT40 cell lines. Our result showed that PL sensitized the cells deficient in homologous recombination related genes. The increase in frequency of the chromosome breaks and the accumulation of the Rad51 protein to chromatin were observed. It indicates that PL is a clastogenic and cytotoxic chemical that induces DSBs. TPZ is converted to a radical intermediate in hypoxia and this intermediate interacts with intracellular macromolecules including DNA. tdp1, tdp2, parp1, and aptx1 cells displayed hyper-sensitivity to TPZ only in hypoxic conditions. These suggest that the accumulations of topo I or topo II-trapped DNA complex and abortive ligation products with 3'-AMP are the potential cause of TPZ-induced killing of hypoxic cells. We conclude that TPZ induces oxidative DNA damage both in normoxia and hypoxia, and introduces abortive topo-DNA complexes and unligatable DNA ends selectively in hypoxia.

Academic Significance and Societal Importance of the Research Achievements

腫瘍細胞特異的な致死効果を持つピペロングミンの解析結果は、正常細胞への影響を最小限に抑える新規抗がん剤の開発に繋がることが期待される。また、腫瘍細胞特有の微細環境（低グルコースや低酸素）に着目し、その環境下でのみ細胞致死効果が得られる薬剤の探索も同様の意義を持つと考える。低酸素下で特異的な致死効果をもたらすチラパザミンが、細胞内活性酸素種のアンバランスによりDNA損傷を誘発するとの結果は、腫瘍細胞内微細環境を利用した新規抗がん剤のスクリーニングにヒントを与えるものである。

Research Products

• [Journal Article] The Effect of p53 Status on Radio-Sensitivity of Quiescent Tumor Cell Population Irradiated With γ-Rays at Various Dose Rates. (2018)
  • Author(s): Masunaga S, Kobayashi J, Tano K, Sanada Y, Suzuki M, Ono K.
  • Journal Title: J Clin Med Res. Volume: 10 Issue: 11 Pages: 815-821
  • DOI: 10.14740/jocmr3610w
  • Peer Reviewed / Open Access
• [Journal Article] Selective cytotoxicity of the anti-diabetic drug, metformin, in glucose-deprived chicken DT40 cells (2017)
  • Author(s): Kadoda K, Moriwaki T, Tsuda M, Sasanuma H, Ishiai M, Takata M, Ide H, Masunaga S, Takeda S and Tano K
  • Journal Title: PLOS ONE Volume: 12 Issue: 9 Pages: e0185141-e0185141
  • DOI: 10.1371/journal.pone.0185141
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effect of Tirapazamine, Metformin or Mild Hyperthermia on Recovery from Radiation-Induced Damage in Pimonidazole-Unlabeled Quiescent Tumor Cells (2017)
  • Author(s): Masunaga S, Tano K, Sanada Y, Sakurai Y, Tanaka H, Suzuki M, Kondo N, Watanabe T, Takata T, Maruhashi A and Ono K
  • Journal Title: World J. Oncol. Volume: 8 Issue: 5 Pages: 137-146
  • DOI: 10.14740/wjon1058w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Complementation of aprataxin deficiency by base excision repair enzymes in mitochondrial extracts (2017)
  • Author(s): Caglayan M, Prasad R, Krasich R, Longley MJ, Kadoda K, Tsuda M, Sasanuma H, Takeda S, Tano K, Copeland WC and Wilson SH
  • Journal Title: Nucleic Acids Res. Volume: 45 Issue: 17 Pages: 10079-10088
  • DOI: 10.1093/nar/gkx654
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cytotoxicity of tirapazamine (3-amino-1, 2, 4- benzotriazine-1, 4-dioxide)-induced DNA damage in chicken DT40 cells. (2017)
  • Author(s): Moriwaki T, Okamoto S, Sasanuma H, Nagasawa H, Takeda S, Masunaga S, Tano K
  • Journal Title: Chem Res Toxicol. Volume: 30 Issue: 2 Pages: 699-704
  • DOI: 10.1021/acs.chemrestox.6b00417
  • Peer Reviewed / Open Access
• [Journal Article] Effect of oxygen pressure during incubation with a 10B-carrier on 10B uptake capacity of cultured p53 wild-type and mutated tumor cells: dependency on p53 status of tumor cells and types of 10B-carriers (2016)
  • Author(s): Masunaga S, Tatebe H, Nishimura Y, Tano K, Sanada Y, Moriwaki T, Sakurai Y, Tanaka H, Suzuki M, Kondo N, Maruhashi A and Ono K
  • Journal Title: International Journal of Radiation Biology Volume: 92 Issue: 4 Pages: 187-194
  • DOI: 10.3109/09553002.2016.1137104
  • Peer Reviewed / Open Access
• [Journal Article] The effect of p53 status of tumor cells on radiosensitivity of irradiated tumors with carbon-ion beams compared with γ-rays or reactor neutron beams (2015)
  • Author(s): Masunaga S, Uzawa A, Hirayama R, Matsumoto Y, Sakurai Y, Tanaka H, Tano K, Sanada Y, Suzuki M, Maruhashi A and Ono K
  • Journal Title: World J. Oncol. Volume: 6 Issue: 4 Pages: 398-409
  • DOI: 10.14740/wjon941w
  • NAID: 120006644056
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 脊椎動物細胞におけるDNA-タンパク質クロスリンク損傷修復へのTDP1, TDP2遺伝子産物の関与 (2018)
  • Author(s): 藤池春奈、Mahmoud Shoulkamy、Amir Salem、津田雅貴、笹沼博之、増永慎一郎、武田俊一、井出博、田野恵三
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] DNAタンパククロスリンク損傷修復におけるFANC及びTDP1遺伝子産物の関与 (2017)
  • Author(s): 藤池 春奈, Mahmoud Shoulkamy, Amir Salem, 笹沼 博之, 高田 穣, 武田 俊一, 増永 慎一郎, 井 出 博, 田野 恵三
  • Organizer: 日本環境変異原学会第46回大会
• [Presentation] DNAタンパククロスリンク損傷修復におけるFANC及びTDP1遺伝子産物の関与 (2017)
  • Author(s): 藤池 春奈, Mahmoud Shoulkamy, Amir Salem, 笹沼 博之, 高田 穣, 武田 俊一, 増永 慎一郎, 井 出 博, 田野 恵三
  • Organizer: 第40回日本分子生物学会年会
• [Presentation] DT40ノックアウト細胞を用いたBiguanide系薬剤Metforminによるグルコース枯渇下細胞致死作用の解析 (2016)
  • Author(s): 角田圭，森脇隆仁，藤池春奈，津田雅貴，笹沼博之，高田穣，井出博，武田俊一，増永慎一郎，田野恵三
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県、横浜市）
  • Year and Date: 2016-11-30
• [Presentation] 低グルコースにおいて致死作用をもつメトフォルミンとフェンフォルミンの作用機序の解析 (2016)
  • Author(s): 角田圭，森脇隆仁，藤池春奈，津田雅貴，笹沼博之，高田穣，井出博，武田俊一，増永慎一郎，田野恵三
  • Organizer: 日本環境変異原学会第45回大会
  • Place of Presentation: つくば国際会議場（茨城県、つくば市）
  • Year and Date: 2016-11-17
• [Presentation] Anti-diabetic biguanide drugs, metformin and phenformin have genotoxicity upon glucose deprivation in chicken DT40 cells (2016)
  • Author(s): Kadoda K, Moriwaki T, Fujiike H, Tsuda M, Sasanuma H, Takata M, Takeda S, Masunaga S, Tano K
  • Organizer: The 10th 3R Symposium
  • Place of Presentation: Hotel Ichihata, Matsue, Japan
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Presentation] DT40ノックアウト細胞パネルを用いたグルコース枯渇下で細胞致死性作用を持つBiguanide薬剤の作用機序の解析 (2016)
  • Author(s): 角田圭，森脇隆仁，藤池春奈，津田雅貴，笹沼博之，高田穣，武田俊一，増永慎一郎，田野恵三
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: ＪＭＳアステールプラザ（広島県、広島市）
  • Year and Date: 2016-10-26
• [Presentation] DT40ノックアウト細胞パネルを用いたグルコース枯渇下で細胞致死作用を持つBiguanide２種の作用機序の解析 (2015)
  • Author(s): 角田圭，森脇隆仁，津田雅貴，笹沼博之，武田俊一，増永慎一郎，田野恵三
  • Organizer: 日本環境変異原学会第４４回大会
  • Place of Presentation: 九州大学馬出キャンパス コラボ・ステーションⅠ・Ⅱ（福岡県、福岡市）
  • Year and Date: 2015-11-27
• [Presentation] DT40細胞のDNA修復欠損細胞パネルを用いた酸素濃度に依存したチラパラザミンのDNA損傷誘発と細胞致死効果の解析 (2015)
  • Author(s): 森脇隆仁，岡本紗季，笹沼博之，永澤秀子，武田俊一，増永慎一郎，田野恵三
  • Organizer: 第68回日本酸化ストレス学会学術集会
  • Place of Presentation: かごしま県民交流センター（鹿児島県、鹿児島市）
  • Year and Date: 2015-06-11
• [Presentation] Characteristics of differential cytotoxicity profile of tirapazamine between normoxia and hypoxia. (2015)
  • Author(s): Moriwaki T, Okamoto S, Sasanuma H, Nagasawa H, Takeda S, Masunaga S and Tano K
  • Organizer: 15th International Congress of Radiation Research (ICRR)
  • Place of Presentation: Kyoto International Conference Center, Kyoto, Japan
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Remarks] 研究室ホームページ
  • URL: http://www.rri.kyoto-u.ac.jp/rb-rri/index.html
• [Remarks] 京都大学原子炉実験所 放射線生命医科学研究本部 放射線生命科学研究部門 粒子線生物学研究分野
  • URL: http://www.rri.kyoto-u.ac.jp/rb-rri/index.html
• [Remarks] 京大原子炉実験所 放射線生命医科学研究本部 放射線生命科学研究部門 粒子線生物学研究分野
  • URL: http://www.rri.kyoto-u.ac.jp/rb-rri/index.html