2000 Fiscal Year Annual Research Report

分子シャペロンを用いたがん治療の基礎的研究

Research Project

Project/Area Number	12670899
Research Institution	Nara Medical University
Principal Investigator	大西健奈良県立医科大学, 医学部, 助教授 (50152195)
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】変異型p53癌細胞に効率のよい癌治療効果をもたらすため、変性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タンパク質に正常型p53タンパク質の正常機能を回復させることを目的とした。【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放射線、温熱あるいは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DNA ladder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Gel mobility-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。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、ヌードマウス移植系ではCDDPによる腫瘍増殖抑制がグリセロール処理で増強された。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p53 consensus sequenceに結合するようになった。【考察】グリセロールを介した変異型p53蛋白質の活性回復により、X線、温熱あるいはCDDP処理後にp53依存的アポトーシスが誘導され、感受性が増強されることが示唆された。【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放射線、温熱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 camptothecin 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

分子シャペロンを用いたがん治療の基礎的研究

Principal Investigator

大西 健 奈良県立医科大学, 医学部, 助教授 (50152195)

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 camptothecin 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)

大西健奈良県立医科大学, 医学部, 助教授 (50152195)

[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)