The Basic Study of Multidisciplinary Treatment for Malignant Tumors using MnSOD Gene Therapy

MnSOD基因治疗多学科综合治疗恶性肿瘤的基础研究

• 批准号: 10470196
• 负责人: KURODA Masahiro
• 金额: $ 2.56万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470196/
• 关键词: MnSOD; gene therapy; radiotherapy; hyperthermia; chemotherapy; tumorigenicity; apoptosis; multidisciplinary treatment; MnSOD; 科学療法

1) We confirmed that MnSOD gene overexpression increased the in vivo effects of radiotherapy, hyperthermia, and chemotherapy, using MnSOD gene-tranfected fibrosarcoma cell line, FSa-II.2) Experiments using scid mice revealed that overexpression of MnSOD gene suppressed spontaneous apoptosis without a resultant alteration in in vivo growth of the mouse fibrosarcoma, FSa-II.3) Newly developed microscopic system was available to indicate that MnSOD gene overexpression increased apoptosis after hyperthermic treatments.4) In vitro timelapse microscopic observation of cellular moving ability did not show any difference by MnSOD gene overexpression.5) Intracellular MnSOD activity following gene-transfection by in vivo electroporation increased as concentration of DNA increased from 5 to 5Oμg/6μl, and reached peak at third day after transfection.6) MnSOD gene-transfection by in vivo electroporation increased the in vivo effects of radiotherapy and hyperthermia. MnSOD gene was transfected by in vivo electroporation for FSa-II tumor in C3H/He mice. Tumors were irradiated at graded doses or heated at 44℃ for 60 min at third day after gene-transfection. The effects of treatments were analyzed by tumor growth-50 method and tumor control dose-50 method.This study indicated that MnSOD overexpression increased thermosensitivity of cells in vitro, and substantially reduces the tumorigenicity, resulting in a significant increase in tumor growth time in vivo. Reduced tumorigenicity by MnSOD overexpression also resulted in a significant increase in tumor control rate by radiotherapy in vivo.

1)我们使用MnSOD基因转染的纤维肉瘤细胞系FSa-II证实MnSOD基因过表达增加了放射治疗、热疗和化学治疗的体内效应。2）使用scid小鼠的实验显示MnSOD基因过表达抑制了小鼠纤维肉瘤的自发凋亡，而不导致体内生长的改变，FSa-II.3）新开发的显微系统可用于表明MnSOD基因过表达增加了热处理后的细胞凋亡。4）在体外时间推移显微镜观察中，MnSOD基因过表达对细胞运动能力没有任何影响。5）在5 ~ 50 μg/6μl的DNA浓度范围内，MnSOD活性随DNA浓度的增加而增加，并在转染后第3天达到峰值。6）MnSOD基因的体内转染增强了放疗和热疗的体内效应。采用电穿孔法将MnSOD基因转染C3 H/He小鼠FSa-II肿瘤。基因转染后第3天，采用不同剂量照射或44℃加热60 min。采用肿瘤生长-50法和肿瘤对照剂量-50法分析治疗效果，结果表明MnSOD过表达增加了体外培养细胞的温度敏感性，并显著降低了致瘤性，导致肿瘤在体内的生长时间显著增加。MnSOD过表达降低致瘤性也导致体内放疗肿瘤控制率显著增加。

项目成果

Osamu Honda et al.: "Assessment of Secondary Necrosis of Jurkat Cells Using a New Microscopic System and Double Staining Method with Annexin V and Propidium Iodide"Int J Oncology. 16. 283-288 (2000)

Osamu Honda 等人：“使用新的显微系统和膜联蛋白 V 和碘化丙啶双重染色方法评估 Jurkat 细胞的继发性坏死”Int J Oncology。

Masahiro Kuroda et al.: "Overexpression of manganese superoxide dismutase gene suppresses spontaneous apoptosis without a resultant alteration in in vivo growth of the mouse fibrosarcoma. FSa-II"Anticancer Res. 7-10 (2000)

Masahiro Kuroda 等人：“锰超氧化物歧化酶基因的过度表达可抑制自发性细胞凋亡，但不会改变小鼠纤维肉瘤的体内生长。FSa-II”Anticancer Res。

Masahiro Kuroda: "A new experimental system for irradiating tumors in mice using a linear accelerator under specific pathogen-free conditions" Acta Med Okayama. 53,2 (in press). (1999)

Masahiro Kuroda：“一种在特定无病原体条件下使用直线加速器照射小鼠肿瘤的新实验系统”Acta Med Okama。

Yoshitomo Ando et al.: "The Effect of Calcium on Fas-Mediated Apoptosis and Secondary Necrosis of Jurkat Cells"Int J Mol Med. 7. 243-247 (2000)

Yoshitomo Ando 等：“钙对 Jurkat 细胞 Fas 介导的细胞凋亡和继发性坏死的影响”Int J Mol Med。

Masahiro Kuroda et al.: "Overexpression of manganese superoxide dismutase gene suppresses spontaneous apoptosis without a resultant alteration in vivo growth of the mouse fibrosarcoma, FSa-II"Anticancer Res. (In press). (2000)

Masahiro Kuroda 等人：“锰超氧化物歧化酶基因的过度表达抑制自发性细胞凋亡，而不改变小鼠纤维肉瘤 FSa-II 的体内生长”Anticancer Res。