Basic Research on gene transfer using gene-gun against malignant gliomas

基因枪抗恶性胶质瘤的基因转移基础研究

• 批准号: 10470285
• 负责人: ASAI Akio
• 金额: $ 7.94万
• 依托单位: The University of Tokyo
• 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 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470285/
• 关键词: gene therapy; gene-gun; glioma; adenovirus

(1) Ex vivo gene transfer with gene-gunWe evaluated gene-gun-mediated ex vivo gene transfer using subcuateous tumor-bearing rat model. First we transplanted 9L glioma cells into F344 syngeneic rats subcutaneously and surgically removed formed tumors 20 days after transplantation. Lac Z expression vector was transduced by gene-guun at shooting pressure (psi) 100, 200, 400, and 600 (max) or Lca Z expressing adenovirus was incubated with the tumor at MOI 20, 40, and 100 for 24 hours. The tumors were then re-transplanted subcutaneously and gene expression was evaluated 7 days after the re-transplantation. The highest gene-gun-mediated gene transfer rate ranged from 3 to 9 % along with shooting pressure whereas that by adenovirus ranged from 5.5 to 15.9% along with MOI which are statistically different.(2) In vivo gene transfer with gene-gunNext, we estimated gene-gun mediated in vivo gene transfer using the same animal model as (1). We surgically opened the skin over the subcutaneously formed tumors and shot Lac Z expression vector at the same psi as (1) or injected Lac Z expression adenovirus at the same MOIs as (1). After the treatments, the wounds were closed and gene expression was examined 7 days after the treatments. The highest gene-gun-mediated gene transfer rate ranged from 3.6 to 9.5 % along with shooting pressure whereas that by adenovirus ranged from 5.8 to 13.2 % along with MOI which are statistically different.In conclusion, although gene transfer with gene-gun is less effective than adenovirus-mediated gene transfer against gliomas, gene-gun-mediated gene transfer is still a good tool to transduce genes in vivo or ex vivo.

(1)基因枪介导的体外基因转移采用皮下荷瘤大鼠模型，对基因枪介导的体外基因转移进行了评价。首先将9L胶质瘤细胞皮下移植到F344同基因大鼠体内，移植20天后手术切除形成的肿瘤。用基因枪在射击压力（psi） 100、200、400和600 （max）下转导Lca Z表达载体，或将表达Lca Z的腺病毒与肿瘤在MOI 20、40和100下孵育24小时。再次皮下移植肿瘤，7天后评估基因表达。基因枪介导的最高基因转移率随射击压力的变化在3% ~ 9%之间，而腺病毒介导的最高基因转移率随MOI的变化在5.5% ~ 15.9%之间，差异有统计学意义。(2)利用基因枪进行体内基因转移。接下来，我们使用与(1)相同的动物模型估计基因枪介导的体内基因转移。我们通过手术打开皮下形成肿瘤的皮肤，以与(1)相同的psi注射Lac Z表达载体，或以与(1)相同的MOIs注射Lac Z表达腺病毒。治疗后缝合创面，于治疗后第7天检测基因表达。基因枪介导的基因转移率随射击压力的变化在3.6% ~ 9.5%之间，腺病毒介导的基因转移率随MOI的变化在5.8% ~ 13.2%之间，差异有统计学意义。综上所述，尽管基因枪介导的基因转移治疗胶质瘤的效果不如腺病毒介导的基因转移，但基因枪介导的基因转移仍然是体内或体外转导基因的良好工具。

项目成果

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Shinoura N、Yoshida Y、Asai A、Kirino T、Hamada H：“Bax 和 Bcl-XiiD2LieD2 的相对表达水平决定了 Bax 过度表达诱导的细胞凋亡/坏死的命运。”Oncogene。18。5703-5713（ 1999）

Ueki K,Chen W-B,Narita Y,Asai A、Kirino T: "Tight association of loss of merlin expression with loss of heterozygosity at chemosome 22q in spoardic meningiomas"Cancer Res. 59. 5995-5998 (1999)

Ueki K、Chen W-B、Narita Y、Asai A、Kirino T：“散发性脑膜瘤中 merlin 表达缺失与化学体 22q 杂合性缺失的紧密关联”Cancer Res. 59. 5995-5998 (1999)

Sano T, Asai A, Mishima K, Fujimaki T, Kirino T: "Telomerase activity in 144 brain tumors"Br J Cancer. 77. 1633-1637 (1998)

Sano T、Asai A、Mishima K、Fujimaki T、Kirino T：“144 种脑肿瘤中的端粒酶活性”Br J Cancer。

Shinoura N., Yoshida Y., Nishimura M., Muramatsu Y., Asai A., Kirino T., Hamada H.: "Expression level of Bcl-2 determines anti- or proapoptotic function"Cancer Res. 59. 4119-4128 (1999)

Shinoura N.、Yoshida Y.、Nishimura M.、Muramatsu Y.、Asai A.、Kirino T.、Hamada H.：“Bcl-2 的表达水平决定抗凋亡或促凋亡功能”Cancer Res。

Chi S, Kitanaka C, Noguchi K, Mochizuki T, Nagashima Y, Shirouzu M, Fujita H, Yoshida M, Chen W, Asai A, Himeno M, Yokoyama S, Kuchino Y: "Oncogenic Ras trigger cell suicide through the activation of a caspase-independent cell death program in human cance

Chi S、Kitanaka C、Noguchi K、Mochizuki T、Nagashima Y、Shirouzu M、Fujita H、Yoshida M、Chen W、Asai A、Himeno M、Yokoyama S、Kuchino Y：“致癌 Ras 通过激活