2000 Fiscal Year Annual Research Report

温熱療法における分子シャペロン・遺伝子治療の応用の基礎的研究

Research Project

Project/Area Number	12217130
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (C)
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)
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 / 遺伝子導入 / p53
Research Abstract	【目的】本研究では、変性した蛋白質構造を正常な蛋白質構造に戻す働きのある分子シャペロン(グリセロール)を用い、変異型p53蛋白質に正常型p53蛋白質の正常機能を回復させることにより、効率のよい温熱癌治療の開発を目指した。【方法】神経膠芽腫細胞A-172あるいは舌扁平上皮癌細胞SASに変異型p53遺伝子(codon248,Arg→Trp)を導入した細胞(A-172/Trp248,SAS/Trp248)および変異型p53を保有する甲状腺未分化癌細胞8305cを用いた。グリセロール処理は温熱、放射線あるいはCDDP処理の2日前に培地に添加し(最終濃度0.6M)、これらストレス処理はグリセロール存在下で行った。コロニー形成法で感受性を測定し、ヘキスト染色法及びDNAladder検出法でアポトーシスの解析を行った。変異型p53蛋白質のDNA結合活性はGelmobility-shift assay法で検討した。【結果】グリセロールによりA-172/Trp248細胞あるいはSAS/Trp248細胞の温熱感受性あるX線感受性が増感し、アポトーシスが増強された。同様に、グリセロール処理をした8305c細胞は、X線あるいはCDDPによるアポトーシス誘導の増強が見られた。また、グリセロールおよび温熱処理したA-172/Trp248細胞から抽出した核内p53タンパク質はp53consensus sequenceに結合するようになった。【考察】以上の結果はグリセロールを介した変異型p53蛋白質の活性回復が起こり、温熱、X線あるいはCDDPによりp53依存的アポトーシスが誘導され、、感受性が増強されたことを示唆している。【結論】変異型p53を保有する癌患者に対しては分子シャペロン治療を併用した温熱、放射線あるいは抗癌剤治療が有効であることを示唆している。

Research Products

(19 results)

All Other

All Publications (19 results)

[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)

2000 Fiscal Year Annual Research Report

温熱療法における分子シャペロン・遺伝子治療の応用の基礎的研究

Principal Investigator

大西 武雄 奈良県立医科大学, 医学部, 教授 (60094554)

Research Products

[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)

大西武雄奈良県立医科大学, 医学部, 教授 (60094554)

[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)