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Hypoxia-to-reoxygenation condition as one of the internal carcinogenic factors of common cancers

缺氧复氧状态是常见癌症的内在致癌因素之一

基本信息

批准号：
17590334
负责人：
OKADA Futoshi
金额：
$ 2.11万
依托单位：
Yamagata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590334/
关键词：
Hypoxia reperfusion Free radical Carcinogenesis DNA microarray Proteomics 悪性化

项目摘要

We examined whether free radicals generated in a series of ischemia-reperfusion condition could act as an endogenous carcinogenic factor in normal or premalignant cells. We obtained various cell lines from human epithelial cells, rodent immortalized fibroblasts and benign fibrosarcoma cells to examine whether those cell lines could be converted into tumorigenic ones after cultivation in a series of hypoxia-reoxygenation condition. Free radicals were produced in the cells after they were cultured in the hypoxia-reoxygenation condition. Peroxides, a part of free radicals, were detected by using the specific probe 2', 7'-dichlorodihydrofluorescein diacetate (H2DCFDA). The production of peroxides was suppressed by an addition of several free radical scavengers into the culture medium. Among those cell lines, non-tumorigenic mouse fibroblast cell lines were converted into tumorigenic ones in nude mice. We then compared their gene/protein expressions with their normal counterparts. We have identified two genes possibly related with carcinogenesis by combinational analyses of DNA microarray and two-dimensional protein electrophoresis. The expression of the two genes decreased in the tumorigenic cells compared to parent cells. We found that one of the two genes actually regulated tumorigenic potential of the mouse fibroblast cells. We expect to further confirm this finding after examining expressions of those genes in a variety of human tumor tissues and rodent spontaneous arising tumors.

我们研究了一系列缺血-再灌注条件下产生的自由基是否可以作为正常细胞或癌前细胞的内源性致癌因子。我们从人上皮细胞、啮齿动物永生化成纤维细胞和良性纤维肉瘤细胞中获得多种细胞系，观察这些细胞系在一系列缺氧-复氧条件下培养后能否转化为致瘤细胞。细胞在缺氧-复氧条件下培养后产生自由基。过氧化氢是自由基的一部分，用特异性探针2',7'-二氯双氢荧光素（H2DCFDA）检测。在培养基中加入几种自由基清除剂可抑制过氧化物的产生。其中，非致瘤性小鼠成纤维细胞系在裸鼠体内转化为致瘤性小鼠。然后，我们将他们的基因/蛋白质表达与正常对照组进行比较。我们通过DNA微阵列和二维蛋白电泳的组合分析鉴定了两个可能与致癌有关的基因。与亲本细胞相比，这两个基因在致瘤细胞中的表达降低。我们发现这两个基因中的一个实际上调控了小鼠成纤维细胞的致瘤潜能。我们希望在检测这些基因在各种人类肿瘤组织和啮齿动物自发肿瘤中的表达后进一步证实这一发现。