ANTI-TUMOR EFFECT OF ANTI-ANGIOGENIC FACTOR GENES PRODUCED BY ELECTROPORATION

电穿孔产生的抗血管生成因子基因的抗肿瘤作用

• 批准号: 14571176
• 负责人: GUNJI Yoshio
• 金额: $ 2.24万
• 依托单位: CHIBA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571176/
• 关键词: 電気穿孔法; 遺伝子治療; 血管新生抑制遺伝子; 抗腫瘍効果; 電撃化学療法; electroporation; angiostatin; endostatin; immunotherapy; gene therapy

The antitumor efficiency of electrochemotherapy using chemotherapeutic agents and high voltage electric pulse has been reported. This study was done to define the precise nature of the involvement of antitumor immunity in the regression of tumor nodules in electrochemotherapy, and to evaluate the effectiveness of using low voltage electroporation. Colon 26 tumors were eradicated in the mice given an intratumor (i.t.) injection of 500 μg bleomycin followed by treatment with electric fields ranging from 50 to 150V/cm, with complete response rates ranging from 80 to 100%. The mice rejected inoculations of re-challenged colon 26 cells, but not Meth A cells. In the Balb/c nu/nu nude mice, complete regression of the tumor was not seen after electrochemotherapy tinder the same therapeutic conditions that resulted in almost complete cure in the Balb/c mice. Our results suggest that the generation of T-cell-dependent, tumor-specific protective immunity might be involved in the process of tumor … More nodule regression in low-voltage electrochemotherapy.Anti-angiogenic factors are potent endothelial cell growth inhibitors that have been shown to inhibit angiogenesis in vitro and tumor growth in mice. We have demonstrated the synergistic anti-tumor effect of anti-angiogenic genes (mouse angiostatin : pBLAST-mAngio, and mouse endostatin : p-BLAST42-mEndo XV) delivered to tumors by low-voltage electroporation in mouse colon 26 models. A synergistic anti-tumor effect was strongly suggested by in vivo tumor growth kinetics, as well as in survival studies with the mice. RT-PCR confirmed the fragments of each gene were transferred by low-voltage electroporation in the tumor. Decreased microvessel density measurements in tumors also confirmed the efficacy of the synergistic antitumor effect of both genes. Significant growth inhibition was observed in mice treated with a 1:1-proportion of angiostatin and endostatin genes, and the order of the both genes transferred (first the endostatin-gene, followed one week later by the angiostatin-gene) had a profound inhibitory effect on tumor growth. These data suggest that in viva delivery of anti-angiogenic genes with low-voltage electroporation could be a possible therapeutic strategy for established solid tumors when both genes were applied in combination. Less

已有文献报道了用化疗药物和高压电脉冲进行电化学治疗的抗肿瘤效果。本研究的目的是明确抗肿瘤免疫参与电化学治疗肿瘤结节消退的确切性质，并评估使用低电压电穿孔的有效性。结肠26肿瘤在给予肿瘤内（i.t.）注射500 μg博莱霉素，然后用50 - 150 V/cm的电场治疗，完全缓解率为80 - 100%。小鼠拒绝再攻击的结肠26细胞的接种，但不是Meth A细胞。在Balb/c nu/nu裸小鼠中，在导致Balb/c小鼠中几乎完全治愈的相同治疗条件下，在电化学疗法后未观察到肿瘤完全消退。结果提示，肿瘤的发生可能与T细胞依赖的肿瘤特异性保护性免疫的产生有关 ...更多信息 抗血管生成因子是有效的内皮细胞生长抑制剂，已显示其在体外抑制血管生成和在小鼠中抑制肿瘤生长。我们已经在小鼠结肠26模型中证明了通过低压电穿孔递送至肿瘤的抗血管生成基因（小鼠血管抑制素：pBLAST-mAngio和小鼠内皮抑制素：p-BLAST 42-mEndo XV）的协同抗肿瘤作用。体内肿瘤生长动力学以及小鼠存活研究强烈表明了协同抗肿瘤作用。RT-PCR证实，每个基因片段在肿瘤中的低电压电穿孔转移。肿瘤中微血管密度测量值的降低也证实了两种基因协同抗肿瘤作用的功效。在用1：1比例的血管抑制素和内皮抑制素基因处理的小鼠中观察到显著的生长抑制，并且两种基因转移的顺序（首先是内皮抑制素基因，一周后是血管抑制素基因）对肿瘤生长具有显著的抑制作用。这些数据表明，在体内交付的抗血管生成基因与低电压电穿孔可能是一个可能的治疗策略，建立实体瘤时，两个基因的组合应用。少

项目成果

Gunji Y., Yol, S., Hayashi, H., Matsubara, H., Hori, S., Suzuki, T., Ochiai, T.: "Analysis of the experimental liver metastasis from inoculated tumor in the mouse stomach cavity"Sung Today. 32. 142-147 (2002)

Gunji Y.、Yol, S.、Hayashi, H.、Matsubara, H.、Hori, S.、Suzuki, T.、Ochiai, T.：“小鼠胃腔内接种肿瘤的实验性肝转移分析”

Shimada, H., Matsubara, Y., Gunji, Y., Tagawa; M., Ochiai T.: "Gene therapy for esophageal cancer."Res.Adv.In cancer. 2. 45-52 (2002)

岛田，H.，松原，Y.，群治，Y.，田川；

Miyazaki, S., Gunji, Y., et al.: "Possible involvement of antitumor immunity in the mechanism of eradication of colon 26 induced by low-voltage electrochemotherapy with bleomycin"Surgery Today. (in press). (2003)

Miyazaki, S.、Gunji, Y. 等人：“抗肿瘤免疫可能参与博来霉素低压电化学疗法诱导的 26 号结肠根除机制”《今日外科》。

上里昌也, 軍司祥雄 et al.: "大きな腫瘍に対するElectrochemotherapy繰り返し施行の効果"日外会誌. 103. 654-654 (2002)

Masaya Kamisato、Yoshio Gunji 等：“重复电化学疗法对大肿瘤的影响”日本盖亚协会杂志 103. 654-654 (2002)。

Miyazaki, S., Gunji.Y.et al.: "Possible invlvement of antitumor immunity in the mechanism of eradication of colon 26 induced by low-voltage electrochemotherapy with bleomycin."Surg Today. 33. 39-44 (2003)

Miyazaki, S., Gunji.Y.等人：“博来霉素低压电化学疗法诱导的26号结肠根除机制中可能存在抗肿瘤免疫的参与。”Surg Today。