分子シャペロンを併用した放射線・温熱癌治療法の基礎的研究

• Project/Area Number: 12217130
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nara Medical University
• Principal Investigator: 大西 武雄 奈良県立医科大学, 医学部, 教授 (60094554)
• Co-Investigator(Kenkyū-buntansha): 大西 健 奈良県立医科大学, 医学部, 助教授 (50152195) ; 高橋 昭久 奈良県立医科大学, 医学部, 講師 (60275336)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥67,900,000 (Direct Cost: ¥67,900,000); Fiscal Year 2004: ¥15,200,000 (Direct Cost: ¥15,200,000); Fiscal Year 2003: ¥13,700,000 (Direct Cost: ¥13,700,000); Fiscal Year 2002: ¥13,500,000 (Direct Cost: ¥13,500,000); Fiscal Year 2001: ¥13,500,000 (Direct Cost: ¥13,500,000); Fiscal Year 2000: ¥12,000,000 (Direct Cost: ¥12,000,000)
• Keywords: 分子シャペロン / グリセロール / 放射線 / 温熱 / 抗癌剤 / アポトーシス / 癌治療 / p53 / 遺伝子導入

Research Abstract

目的
培養細胞系で確認したグリセロールあるいはp53C末端ペプチド(アミノ酸残基361-382、353-374)の変異型p53タンパク質に対する分子シャペロン効果がヌードマウス移植系でも見られるか検討した。
方法と結果
1.p53ドミナントネガティブの性質を持つ変異型p53導入舌扁平上皮癌細胞SAS/mp53細胞またはneoコントロールSAS/neo細胞(正常型p53表現型)をヌードマウス(Balb/cAJcl-nl)の大腿皮下に移植し、グリセロール局所注入後にX線照射した。グリセロールをあらかじめ注入すると、変異型p53腫瘍において放射線単独の場合よりも明確な増殖遅延効果が見られた。培養細胞を用いた実験と同様にヌードマウス移植癌実験においてもグリセロールの分子シャペロン効果が確認された。
2.グリセロール処理によって放射線誘導アポトーシスが変異型p53腫瘍において増強するかどうかをApoptosis免疫染色によって調べた結果、グリセロール処理によって変異型p53腫瘍でも正常型p53腫瘍と同様の陽性細胞の増加が認められた。
3.p53を欠損した非小細胞肺癌細胞H1299に変異型p53を導入したH1299/mp53細胞(変異型p53)またはneoコントロールベクターのみを導入したH1299/neo細胞(p53欠損)をヌードマウスに移植し、p53C末端ペプチドを局所注入した。変異型p53腫瘍はp53欠損腫瘍に比べ明確な増殖遅延を示した。
考察
以上の結果によって培養細胞を用いた実験と同様にヌードマウス移植癌実験でグリセロールおよびp53C末端ペプチドの分子シャペロン効果が確認された。分子シャペロン治療は変異型p53腫瘍に正常型p53タンパク質の機能を発揮させ、正常型p53タンパク質を持つ腫瘍と同程度の癌治療効果をもたらすことが強く示唆された。

Research Products

• [Journal Article] C-terminal peptides of p53 molecules enhance radiation-induced apoptosis in human mutant p53 cancer cells. (2004)
  • Author(s): Ohnishi K. et al.
  • Journal Title: Apoptosis 9 Pages: 591-597
• [Journal Article] Effects of a heat shock protein inhibitor KNK437 on heat sensitivity and heat tolerance in human squamous cell carcinoma cell lines differing in p53 status alone. (2004)
  • Author(s): Ohnishi K. et al.
  • Journal Title: International Journal of Radiation Biology 80 Pages: 607-614
• [Journal Article] Hyperthermic cancer therapy combined with inhibitors targeted against heat-induced signaling factors. (2004)
  • Author(s): Ohnishi K. et al.
  • Journal Title: Japanese Journal of Hyperthermic Oncology 20 Pages: 143-159
  • NAID: 10029675920
• [Journal Article] Glycerol enhances CDDP-induced growth inhibition of thyroid anaplastic carcinoma tumor carrying mutated p53 gene. (2004)
  • Author(s): Yuki K.et al.
  • Journal Title: Oncology Report 11 Pages: 821-824
• [Journal Article] Sensitization by glycerol for CDDP-therapy against human cultured cancer cells and tumors bearing mutated p53 gene. (2004)
  • Author(s): Yuki K.et al.
  • Journal Title: Apoptosis 9 Pages: 853-859
• [Journal Article] Evidence for the involvement of double-strand breaks in heat-induced cell killing. (2004)
  • Author(s): Takahashi A. et al.
  • Journal Title: Cancer Research 64 Pages: 8839-8845
• [Journal Article] High-LET radiation enhanced apoptosis but not necrosis regardless of p53 status. (2004)
  • Author(s): Takahashi A. et al.
  • Journal Title: International Journal of Radiation Oncology Biology Physics 60 Pages: 591-597
• [Journal Article] Apoptosis-related gene expression after hyperthermia in human tongue squamous cell carcinoma cells harboring wild-type or mutated-type p53 (2004)
  • Author(s): Yasumoto J. et al.
  • Journal Title: Cancer Letter 204 Pages: 41-51
• [Journal Article] Effects of mild temperature hyperthermia and p53 status on the size of hypoxic fractions in sold tumors, with reference to the effect in intratumor quiescent cell populations. (2004)
  • Author(s): Masunaga S. et al.
  • Journal Title: International Journal of Radiation Oncology Biology Physics 60 Pages: 570-577
• [Journal Article] 免疫力の活性化も期待できるガン細胞を熱攻めする治療法 (2004)
  • Author(s): 大西武雄
  • Journal Title: Natural Medicine 4 Pages: 1-11
• [Journal Article] ハイパーサーミアによるがん治療-最近のトピックス
  • Author(s): 大西武雄
  • Journal Title: 放射線医学研究所放射線科学 印刷中
• [Publications] Yuki, K., et al.: "Glycerol enhances CDDP-induced growth inhibition of thyroid anaplastic carcinoma tumor carrying mutated p53 gene."Oncol.Rep.. 11(4). 821-824 (2004)
• [Publications] Yasumoto, J., et al.: "Apoptosis-related gene expression after hyperthermia in human tongue squamous cell carcinoma cells harboring wild-type or mutated p53."Cancer Lett.. 204(1). 41-51 (2004)
• [Publications] Yasumoto, J., et al.: "Analysis of apoptosis-related gene expression after X-ray irradiation in human tongue squamous cell carcinoma cells harboring wild-type or mutation-type p53 gene."J.Rad.Res.. 44(1). 41-45 (2003)
• [Publications] Harima, Y., et al.: "Expression of Ku80 in cervical cancer correlates with response to radiotherapy and survival."Amer.J.Clin.Oncol.. 26(4). e80-e85 (2003)
• [Publications] Masunaga S, et al.: "Usefulness of combined treatment with mild hyperthermia and/or tirapazamine in the treatment of solid tumors : its independence of p53 status."Cancer Sci.. 94(1). 125-133 (2003)
• [Publications] Takahashi, A., et al.: "p53-Dependent hyperthermic enhancement of tumor growth inhibition by X-ray or carbon-ion beam irradiation."Int.J. Hyperthermia. 19(2). 145-153 (2003)
• [Publications] Tamamoto, T., et al.: "Heat-induced growth inhibition and apoptosis in transplanted human head and neck squamous cell carcinomas with different status of p53"Int.J. Hyperthermia. 19(2). 590-597 (2003)
• [Publications] Yasumoto, J., et al.: "The role of p53 molecule in radiation and hyperthermic therapies."J. Health Sci.. 49. 273-277 (2003)
• [Publications] 幸和恵 他: "がん細胞に特徴的なシグナル伝達分子を標的とする薬剤"放射線生物研究. 38(3). 245-259 (2003)
• [Publications] Yasumoto, J., et al.: "Analysis of apoptosis-related gene expression after X-ray irradiation in human tongue squamous cell carcinoma cells harboring wild-type or mutation-type p53 gene"J. Rad. Res.. (in press). (2003)
• [Publications] Harima, Y., et al.: "Expression of Ku80 in cervical cancer correlates with response to radiotherapy and survival"Amer. J. Clin. Oncol.. (in press). (2003)
• [Publications] Masunaga S, et al.: "Usefulness of combined treatment with mild hyperthermia and/or tirapazamine in the treatment of solid tumors : its independence of p53 status"Cancer Science. (in press). (2003)
• [Publications] Takahashi, A., et al.: "p53-Dependent hyperthermic enhancement of tumor growth inhibition by X-ray or carbon-ion beam irradiation"Int. J. Hyperthermia. (in press). (2003)
• [Publications] Takahashi, A., et al.: "Radiation-induced apoptosis in scid mouse spleen after low dose-rate irradiation"Int. J. Radiat. Biol.. 78. 689-693 (2002)
• [Publications] Harima, Y., et al.: "Human papilloma virus (HPV) DNA associated with prognosis of cervical cancer after radiotherapy"Int. J. Radiat. Oncol. Biol. Phys.. 52. 1345-1351 (2002)
• [Publications] Harima, Y., et al.: "Mutation of the PTEN gene in advanced cervical cancer correlated with tumor progression and poor outcome after radiotherapy"Int. J. Oncol.. 18. 493-497 (2002)
• [Publications] Masunaga, S., et al.: "Impact of p53 status of the tumor cells on the effect of reactor neutron beam irradiation with emphasis on the response of intratumor quiescent cells"Jpn. J. Cancer Res.. 93. 1366-1377 (2002)
• [Publications] Ohnishi, K., et al.: "Glycerol as a chemical chaperone enhanced radiation-induced apoptosis in anaplastic thyroid carcinoma cells"Mol. Cancer. 1. No.4 (2002)
• [Publications] Asakawa, I., et al.: "Growth inhibition in transplanted human tongue tumors bearing different p53 gene status with X-ray or C-beam irradiation"Anticancer Res.. 22. 2037-2043 (2002)
• [Publications] Ohnishi, K., et al.: "Transfection of mutant p53 gene depresses X-ray-and CDDP-induced apoptosis in a human squamous cell carcinoma of the head and neck"Apoptosis. 7. 367-372 (2002)
• [Publications] Ohnishi, T., et al.: "Glycerol restores heat-induced p53-dependent apoptosis of human glioblastoma cells bearing mutant p53"BioMed. Biotechnol.. 2. No.6 (2002)
• [Publications] Masunaga, S., et al.: "Radiobiological characteristics of solid tumours depending on the p53 status of the tumour cells, with emphasis on the response of intratumour quiescent cells"Eur. J. Cancer. 18. 718-727 (2002)
• [Publications] 大西健 他: "集学的癌治療の研究と臨床"篠原出版新社. 7 (2002)
• [Publications] 太田一郎 他: "癌放射線療法2002"篠原出版新社. 12 (2002)
• [Publications] 村山千恵子 他: "集学的癌治療の研究と臨床"篠原出版. 9 (2002)
• [Publications] 玉本哲郎 他: "癌の臨床"篠原出版. 9 (2002)
• [Publications] Takahashi, A., et al.: "Radiation-induced apoptosis in scid mouse spleen after low dose-rate irradiation"Int.J.Radiat.Biol.. (in press). (2002)
• [Publications] Harima, Y., et al.: "Human papilloma virus(HPV)DNA associated with prognosis of cervical cancer after radiotherapy"Int.J.Radiat.Oncol.Biol.Phys.. (in press). (2002)
• [Publications] Harima, Y., et al.: "Mutation of the PTEN gene in advanced cervical cancer correlated with tumor progression and poor outcome after radiotherapy"Int.J.Oncol.. 18. 493-497 (2002)
• [Publications] Takahashi, A., et al.: "p53-dependent thermal enhancement of cellular sensitivity in human squamous cell carcinomas in relation to LET"Int.J.Radiat.Biol.. 77. 1043-1051 (2001)
• [Publications] Takahashi, A., et al.: "Radiation response of apoptosis in C57BL/6N mouse spleen after whole-body irradiation"Int.J.Radiat.Biol.. 77. 939-946 (2001)
• [Publications] Harima, Y., et al.: "Chromosome 6p21.2,18q21.2 and human papilloma virus(HPV)DNA can predict prognosis of cervical cancer after radiotherapy"Int.J.Cancer.. 96. 286-296 (2001)
• [Publications] Ohnishi, T., et al.: "Tumor suppressor p53 response is blunted by pre-irradiation at a low-dose-rate radiation"Physica Medica(Eur.J.Med.Physics). 17Suppl.1. 215-216 (2001)
• [Publications] Harima, Y., et al.: "Polymorphism of the WAF1 gene is related to susceptibility to cervical cancer in Japanese women"Int.J.Mol.Medicine. 7. 261-264 (2001)
• [Publications] Ohnishi, K., T.Ohnishi: "Heat-induced p53-dependent signal transduction and its role in hyperthermic cancer therapy"Int.J.Hyperthermia. 17. 415-427 (2001)
• [Publications] Matsumoto, H., et al.: "Induction of radioresistance by a nitric oxide-mediated bystander effect"Radiat.Res.. 155. 387-396 (2001)
• [Publications] Kirita, T., et al.: "A new strategy for cancer therapy based on a predictive indicator"Human Cell. 14. 1-6 (2001)
• [Publications] 大西健 他: "集学的癌治療の研究と臨床"篠原出版新社. 7 (2002)
• [Publications] 太田一郎 他: "癌放射線療法2002"篠原出版新社. 12 (2002)
• [Publications] 桐田忠昭 他: "脳の科学"星和書店. 7 (2001)
• [Publications] Harima,Y., et al.: "Polymorphism of the WAF1 gene is related to susceptibility to cervical cancer in Japanese women."Int.J.Mol.Medicine. (in press). (2001)
• [Publications] Harima,Y., et al.: "Mutation of the PTEN gene in advanced cervical cancer correlated with tumor progression and poor outcome after radiotherapy."Int.J.Oncol.. (in press). (2001)
• [Publications] Matsumoto,H., et al.: "Induction of radioresistance by a nitric oxide-mediated bystander effect."Radiat.Res.. 155. 387-396 (2001)
• [Publications] Ohnishi,K., et al.: "Glycerol restores p53 dependent radiosensitivity of human head and neck cancer cells bearing mutant."Br.J.Cancer. 83. 1735-1739 (2000)
• [Publications] Kubota,N., et al.: "Wortmannin sensitizes human glioblastoma cell lines carrying mutant or wild type TP53 gene to radiation."Cancer Lett.. 161. 141-147 (2000)
• [Publications] Oka,A., et al.: "Determination of p53-mediated transactivationl ability in radiation-treated cervical cancer."Cancer Detect.Prevent.. 24. 275-282 (2000)
• [Publications] Kanata,H., et al.: "CDDP induces p53-dependent apotosis in tongue cancer cells."Int.J.Oncol.. 17. 513-518 (2000)
• [Publications] Xie,X., et al.: "p53 status plays no role in radiosensitizing effects of SN-38, a comptothecin derivative."Cancer Chemother.Pharmacol.. 45. 362-368 (2000)
• [Publications] Ohnishi,T., et al.: "Heat-induced accumulation of p53 and hsp72 is suppressed in lung fibroblasts from SCID mouse."Int.J.Radiat.Biol.. 76. 711-715 (2000)
• [Publications] Ota,I., et al.: "Transfection with mutant p53 gene inhibits heat-induced apoptosis in a head and neck cell line of human squamous cell carcinoma."Int.J.Radiat.Oncol.Biol.Phys.. 47. 495-501 (2000)
• [Publications] Takahashi,A., et al.: "The dependence of p53 on the radiation enhancement of thermosensitivity at different LET."Int.J.Radiat.Oncol.Biol.Phys.. 47. 489-494 (2000)
• [Publications] Takahashi,A., et al.: "Induction of WAF1 accumulation by carbon-ion beam and α-particle irradiation in human glioblastoma cultured cells."Int.J.Radiat.Biol.. 76. 335-341 (2000)
• [Publications] Ohtsubo,T., et al.: "Effect of acidic environment and p53 on apoptosis induction by hyperthermia."Int.J.Hyperthermia. 16. 481-491 (2000)
• [Publications] Matsumoto,H., et al.: "Induction of radioresistance to accelerated carbon-ion beams in recipient cells by nitric oxide excreted from irradiated cells of human glioblastoma."Int.J.Radiat.Biol.. 76. 1649-1657 (2000)
• [Publications] Matsumoto,H., et al.: "Interactive effects of nitric oxide and p53 on cellular thermosensitivity."Jpn.J.Hyperthermic Oncol.. 16. 69-82 (2000)
• [Publications] Harima,Y., et al.: "Loss of heterozygosity on chromosome 6p21.2 as a potential marker for recurrence after radiotherapy of human cervical cancer."Clinical Cancer Res.. 6. 1079-1085 (2000)
• [Publications] Harima,Y., et al.: "Bax and Bcl-2 protein expression following radiation therapy versus radiation plus thermoradiotherapy in stage IIIb cervical carcinoma."Cancer. 88. 131-137 (2000)
• [Publications] 太田一郎 他: "癌放射線療法2000"篠原出版(印刷中). (2001)
• [Publications] 桐田忠昭 他: "脳の科学"星和書店(印刷中). (2001)