STUDIES ON ACTIONS OF TUMOR PROMOTERS BY ANALYSIS OF GENE EXPRESSION IN CELL TRANSFORMATION
通过分析细胞转化中的基因表达来研究肿瘤促进剂的作用
基本信息
- 批准号:10672118
- 负责人:
- 金额:$ 2.11万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 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:
- 关键词:
项目摘要
This study was intended to clarify the molecular process of tumor promotion by finding genes that regulate tumor promotion among genes whose expressions are altered by tumor promoters. A two-stage transformation assay in BALB/3T3 cells was used as a model of two-stage carcinogenicity tests. 12-O-tradecanoylphorbol-13-acetate (TPA), okadaic acid and orthovanadate were found to be potent promoters of cell transformation. The total RNA was extracted from the cells being treated with one of these three chemicals under the same conditions as in the promotion phase of the transformation assay, and the mRNA was visualized by a fluorescent mRNA differential display method. Two cDNA fragments, G09 and C05, were isolated. G09 was the cDNA fragment from the mRNA that was commonly increased by three tumor promoters, and the sequence of G09 was entirely identical to 2198-2708 of cDNA (3488 bp) of Np95, a murine nuclear protein. C05 was the 297 bp fragment from the mRNA that was commonly decreased b … More y the tumor promoters, and any homologous sequence of C05 has not been found in data base presented by GenBank + EMBL + DDBJ + PDB.The altered expressions of these two genes were confirmed by RT-PCR using specific primers and northern blotting. It is reported that Np95 protein appears in the S phase of the cell cycle in normal mouse thymocytes and is irregularly expressed in all the murine lymphoma cells. In BALB/3T3 fibroblasts the DNA synthesis measured by BrdU uptake and the expression of Np95 were elevated by proliferative stimuli of serum, and the DNA synthesis was increased in parallel with the elevation of Np95 expression at various times under the treatment by tumor promoters. These results suggest that the expression of Np95 also associates with cell proliferation in the fibroblasts. The fact that the expression of two genes were commonly altered by tumor promoters suggests that there may exist a common mechanism in tumor promotion by these chemicals, though further investigation is required to find the roles of two genes in tumor promotion. Less
本研究旨在通过在肿瘤启动子改变表达的基因中寻找调节肿瘤促进的基因来阐明肿瘤促进的分子过程。 BALB/3T3 细胞的两阶段转化测定被用作两阶段致癌性测试的模型。 12-O-tradecanoylphorbol-13-acetate (TPA)、冈田酸和原钒酸盐被发现是细胞转化的有效促进剂。在与转化测定的促进阶段相同的条件下,从用这三种化学物质之一处理的细胞中提取总RNA,并通过荧光mRNA差异显示方法使mRNA可视化。分离出两个cDNA片段,G09和C05。 G09是由三个肿瘤启动子共同增加的mRNA的cDNA片段,G09的序列与鼠核蛋白Np95的cDNA(3488bp)的2198-2708完全相同。 C05是mRNA中的297 bp片段,在肿瘤启动子中通常会减少,在GenBank + EMBL + DDBJ + PDB提供的数据库中尚未发现任何与C05同源的序列。使用特异性引物和Northern blotting通过RT-PCR证实了这两个基因的表达改变。据报道,Np95蛋白出现在正常小鼠胸腺细胞的细胞周期S期,并且在所有小鼠淋巴瘤细胞中不规则表达。在BALB/3T3成纤维细胞中,血清的增殖刺激使通过BrdU摄取测量的DNA合成和Np95的表达升高,并且在肿瘤促进剂的处理下,不同时间的DNA合成与Np95表达的升高平行地增加。这些结果表明 Np95 的表达也与成纤维细胞中的细胞增殖相关。两个基因的表达通常被肿瘤启动子改变的事实表明,这些化学物质在肿瘤促进中可能存在共同的机制,尽管需要进一步研究来发现这两个基因在肿瘤促进中的作用。较少的
项目成果
期刊论文数量(20)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Ayako Sakai: "p-Nonylphenol acts as a promoter in the BALB/3T3 cell transformation"Mutation Research. (in press). (2001)
Ayako Sakai:“对壬基酚在 BALB/3T3 细胞转化中充当启动子”突变研究。
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- 影响因子:0
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- 通讯作者:
Sakurai, H., Umeda, M., Tsuchiya, T., Sakai, A., Shishiyama, A., et al.: "Japanese Standards Association"TR T 0000 : 2001 : A short-term two-stage cell transformation assay using BALB/c 3T3 cells for the prediction of carcinogenic potential of chemicals (
Sakurai, H.、Umeda, M.、Tsuchiya, T.、Sakai, A.、Shishiyama, A.等:“日本标准协会”TR T 0000:2001:短期两阶段细胞转化测定
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- 影响因子:0
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Ayako Sakai,Reiko Teshima: "2,5-Di-tert-butyl-1,4-hydroquinone enhances cell transformation accompanying an increase in intracellular free calcium ion concentration"Cancer Letters. (in press). (2001)
Ayako Sakai、Reiko Teshima:“2,5-二叔丁基-1,4-氢醌可增强细胞转化,同时增加细胞内游离钙离子浓度”《癌症快报》。
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- 影响因子:0
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Ayako Sakai,Yoko Yamakoshi,Naoki Miyata: "Visible light irradiation of [60] fullerene causes killing and initiation of transformation in BALB/3T3 cells"Fullerene Science and Technology. 7(5). 743-756 (1999)
Ayako Sakai、Yoko Yamakoshi、Naoki Miyata:“[60]富勒烯的可见光照射导致 BALB/3T3 细胞死亡并引发转化”富勒烯科学与技术。
- DOI:
- 发表时间:
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- 影响因子:0
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- 通讯作者:
Sakai, A.and Teshima, R.: "2,5-Di-tert-butyl-1, 4-hydroquinone enhances cell transformation accompanying an increase in intracellular free calcium ion concentration."Cancer Letters. (in press). (2001)
Sakai, A. 和 Teshima, R.:“2,5-二叔丁基-1, 4-氢醌可增强细胞转化,同时增加细胞内游离钙离子浓度。”《癌症快报》。
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SAKAI Ayako其他文献
SAKAI Ayako的其他文献
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{{ truncateString('SAKAI Ayako', 18)}}的其他基金
ANALYSIS OF ALTERED GENE EXPRESSIONS BY TUMOR PROMOTERS DURING CELL TRANSFORMATION
细胞转化过程中肿瘤启动子改变的基因表达的分析
- 批准号:
14572120 - 财政年份:2002
- 资助金额:
$ 2.11万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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