2002 Fiscal Year Annual Research Report

口腔癌に対する遺伝子治療の放射線・化学・温熱療法への応用に関する基礎的研究

Research Project

Project/Area Number	13672110
Research Institution	Nara Medical University
Principal Investigator	桐田忠昭奈良県立医科大学, 医学部, 教授 (70201465)
Co-Investigator(Kenkyū-buntansha)	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 / p53
Research Abstract	【目的】変性した蛋白質構造を正常な蛋白質構造に戻す働きがあるとされる分子シャペロン(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、効率のよい放射線による癌治療の開発を目指した。【方法】変異型p53を持つヒト口腔扁平上皮癌細胞株Ca9-22(codon 248,Arg-Trp)を用いた。グリセロール処理は放射線および温熱処理の2日前に培地に添加した(最終濃度0.6M)。コロニー形成法でX線感受性を測定し、アポトーシス関連タンパク質の定量をWestern Blot法で、アポトーシス誘導をヘキスト染色法及びDNA ladder検出法にてp53のDNA結合量をゲルシフトアッセイにて解析した。【結果】1.グリセロールによりCa9-22細胞のX線感受性が増感を示すようになった。2.X線単独照射の場合、Baxの蓄積誘導はほとんど見られなかった。3.グリセロール処理によりX線によるBaxの蓄積誘導が見られるようになった。4.X線単独照射の場合、DNAラダーは見られないがグリセロール処理を併用した場合、DNAラダーが認められた。温熱の場合はグリセロールによる感受性の増強効果は見られなかった。5.グリセロールとX線の併用により、p53のDNA結合量の増加が見られた。【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線処理後にBaxを介した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治療が有効であることが培養細胞系で示唆された。

Research Products
(20 results)

All Other

All Publications (20 results)

[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] Ohnishi, K., et al.: "P53-dependent hyperthermic enhancement of tumour inhibition by Xray or Carbon-ion beam irradiation"Int. J. Hyperthermia. Mol. Cancer.. (in press). (2003)
[Publications] Masunaga, S,. et al.: "Usefulness of combined treatment with mild hyperthermia and/or tirapazamine in the treatment of solid -its independence of p53 status"Cancer Science,. (in press). (2003)
[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-or CDDP-induced apoptosis in a human sauamous cell carcinoma of the head and neck"Apoptosis. 7. 367-372 (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] 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] Takahashi, A., et al.: "p53-dependent thermal enhancement of cellular sensitivity in human squamous cel 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] Olinishi, T., et al.: "Tumor suppressor p53 response is blunted by pre-irradiation at a low-dose-rate radiation"Physica Medica (Eur. J. Med. Physics). 17 Suppl.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. Hyperthmnia.. 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"星和出版. 7 (2001)

2002 Fiscal Year Annual Research Report

口腔癌に対する遺伝子治療の放射線・化学・温熱療法への応用に関する基礎的研究

Principal Investigator

桐田 忠昭 奈良県立医科大学, 医学部, 教授 (70201465)

Research Products

[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] Ohnishi, K., et al.: "P53-dependent hyperthermic enhancement of tumour inhibition by Xray or Carbon-ion beam irradiation"Int. J. Hyperthermia. Mol. Cancer.. (in press). (2003)

[Publications] Masunaga, S,. et al.: "Usefulness of combined treatment with mild hyperthermia and/or tirapazamine in the treatment of solid -its independence of p53 status"Cancer Science,. (in press). (2003)

[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-or CDDP-induced apoptosis in a human sauamous cell carcinoma of the head and neck"Apoptosis. 7. 367-372 (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] 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] Takahashi, A., et al.: "p53-dependent thermal enhancement of cellular sensitivity in human squamous cel 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] Olinishi, T., et al.: "Tumor suppressor p53 response is blunted by pre-irradiation at a low-dose-rate radiation"Physica Medica (Eur. J. Med. Physics). 17 Suppl.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. Hyperthmnia.. 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"星和出版. 7 (2001)

桐田忠昭奈良県立医科大学, 医学部, 教授 (70201465)

[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"星和出版. 7 (2001)