Estimation of Biologic and Molecular Aspects of Heavy Charged Particle Therapy for Cancer Cells

重带电粒子治疗癌细胞的生物学和分子方面的评估

• 批准号: 13670938
• 负责人: SUGIMOTO Kouji
• 金额: $ 2.3万
• 依托单位: Kobe University Graduate School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670938/
• 关键词: Radiation Oncology; Particle Therapy; DNA double strand Break (DSB); DNA Repair Mechanisms; Rad51; Comet Assay; Radiosensitivity; Antisense; DNA二重鎖切断; DNA修復; DNA; 併用治療; 細胞周期

Carbon ion irradiation has well-balanced dual actions on cancers : efficient doselocalization due to sharp penumbra and distal taill off in dose, and potent biological anticancer effect due to high relative biological effect (RBE). However, biological and molecular mechanisms of carbon ion irradiation have not been clarified. In this study, we focused on the biologic mechanisms of carbon ion irradiation, especially assessing DNA double strand breaks (DSBs) and repair.Cell-killing effect of X-ray and carbon ion irradiation was assessed using a colony-formation assay. The amount of double strand breaks and repair rate was assessed by comat assay. In the comparison of DSBs repair between carbon ion irradiation and X-ray irradiation, the degree of residual DSBs was severe in carbon ion irradiated cells. The expression level of DSB repair associated protein was assessed by Westem blot analysis. Sub-nuclear localization of DNA repair protein Rad51 were immunohistochemically evaluated via epi … More tluorescence microscopy. Rad51 nuclear foci formation was observed after X-ray irradiation and carbon ion irradiation. These nuclear foci in the cells treated with carbon ion were sustained to 24 hours after irradiation, whereas in the cells treated with X-ray irradiation more than half of nuclear foci were disappeared at 24 hours after irradiation. Rad51 antisense oligonucleotides was transfected and irradiated with carbonions. Clonogenic assay after transfection revealed that Rad51 antisense oligonucleotides increased radiosensitivity very little carbon ion irradiation, whereas ti increased about 1.7 fold in X-irradiation. Comet assay was also performed for quantitative analysis of DSB induction and repair. There was no difference of DSB induction and repair rate after carbon ion irradiation in the cells treated with Rad51 antisense oligonucleotides and with control oligos. These results may suggest that carbon-ion-induced DSBs are difficult to be repaired by homologous recombination. Less

碳离子照射对癌症有很好的双重作用：由于尖峰的半影区和远端剂量的下降而有效的剂量定位，以及由于高的相对生物效应(RBE)而产生的强大的生物抗癌效应。然而，碳离子辐照的生物学和分子机制尚不清楚。在本研究中，我们重点研究了碳离子辐射的生物学机制，特别是DNA双链断裂及其修复。采用集落形成实验评价了X射线和碳离子辐射对细胞的杀伤作用。用彗星试验测定双链断裂数和修复率。在碳离子照射和X射线照射的DSB修复的比较中，碳离子照射的细胞中残留的DSB程度较重。用免疫印迹法检测DSB修复相关蛋白的表达水平。用免疫组织化学EPI…法检测DNA修复蛋白RAD51的亚核定位更多的荧光显微镜。X射线照射和碳离子照射后，观察到RAD51核团的形成。碳离子处理组的这些核灶在照射后24小时内持续存在，而X射线照射组在照射后24小时，超过一半的核灶消失。用碳离子照射RAD51反义寡核苷酸。克隆形成实验表明，RAD51反义寡核苷酸对碳离子辐射的增敏作用很小，而对X射线辐射的增敏作用约为后者的1.7倍。彗星试验定量分析DSB的诱导和修复。RAD51反义寡核苷酸组和对照组细胞经碳离子照射后，DSB的诱导率和修复率无明显差异。这些结果可能表明，碳离子诱导的DSB很难通过同源重组来修复。较少

项目成果

Sasaki R: "Angiosarcoma treated with radiotherapy : impact of tumor type and size on outcome"Int J Radiat Oncol Biol Phys. 54(2). (2002)

Sasaki R：“用放射疗法治疗血管肉瘤：肿瘤类型和大小对结果的影响”Int J Radiat Oncol Biol Phys。

Kagawa K, Hishikawa Y, et al: "Preclinical biological assessment of proton and carbon ion beams at Hyogo Ion Beam Medical Center"Int Radiat Oncol Biol Phys. 54(3). 928-938 (2002)

Kakawa K、Hishikawa Y 等人：“兵库离子束医疗中心质子和碳离子束的临床前生物学评估”Int Radiat Oncol Biol Phys。

Nishimura H, Sugimura K, et al: "Radiographic pulmonary and pleural changes after carbon ion irradiation"Int J Radiat Oncol Biol Phys. 55(4). 861-866 (2003)

Nishimura H、Sugimura K 等人：“碳离子照射后放射学肺部和胸膜变化”Int J Radiat Oncol Biol Phys。

Kagawa K: "Preclinical biological assessment of proton and carbon ion beams at Hyogo Ion Beam Medical Center"Int J Radiat Oncol Biol Phys. 54(3). 928-938 (2002)

Kakawa K：“兵库离子束医疗中心质子和碳离子束的临床前生物学评估”Int J Radiat Oncol Biol Phys。

Nishimura H: "Radiographic pulmonary and pleural changes after carbon ion irradiation"Int J Radiat Oncol Biol Phys. 55(4). 861-866 (2003)

Nishimura H：“碳离子照射后放射学肺部和胸膜变化”Int J Radiat Oncol Biol Phys。