DNA repair defect, tumorigensis, and hereditary breast cancer gene Brca2 -Studies in vitro and in vivo, and association with Rad51.

DNA 修复缺陷、肿瘤发生和遗传性乳腺癌基因 Brca2 - 体外和体内研究以及与 Rad51 的关联。

• 批准号: 11460133
• 负责人: MORIMATSU Masami
• 金额: $ 8.32万
• 依托单位: Iwate University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11460133/
• 关键词: breast cancer; dog; cat; mouse; Brca2; Rad51; IkB; 標的遺伝子組換え; 遺伝子機能解析

To establish the bases of developing new methods for prevention, diagnosis, and therapy of breast cancer , we investigated in vitro and invivo functions of the Brca2 and other tumor associated genes. Especially, researches for the significance of the interaction of Brca2 and Rad51, application of Brca2 analysis in veterinary medicine, and roles IkB MAIL were carried out.1. Cloning and mutation analysis of canine Brca2Full length (about 10 kbp) of the cDNA for canine Brca2 was cloned and sequenced. For mutation analysis, DNA fragments were obtained by PCR and used for protein truncation tests.2. Cloning and expression of feline Brca2Full length (about 10 kbp) of the cDNA for feline Brca2 was cloned and sequenced. Expression of feline Brca2 was confirmed in mamnary gland.3. Interaction of Brca2 and Rad51Interaction of C-terminus of canine Brca2 with Rad51 was demonstrated by yeast two-hybrid assay. Transactivation activity of canine Brca2 was also demonstrated.4. Characterization of a novel IkB protein, MAILWe cloned, sequenced, and analyzed the expression of the MAIL gene encoding a novel IkB protein.

本研究对BRCA2等肿瘤相关基因的体内外功能进行了研究，为乳腺癌的预防、诊断和治疗提供新的手段。特别是对BRCA2与RAD51相互作用的意义、BRCA2分析在兽医中的应用以及IKB Mail的作用进行了研究。犬Brca2基因的克隆和突变分析克隆了犬Brca2基因的全长(约10kbp)，并对其进行了序列分析。突变检测采用聚合酶链式反应获得DNA片段，并进行蛋白质截短试验。猫Brca2全长(约10kbp)的克隆和表达。证实了猫科动物BRCA2在乳腺组织中的表达。用酵母双杂交实验证实了犬BRCA2的C末端与RAD51的相互作用。还证实了犬BRCA2的反式激活活性。新的IKB蛋白MAIL的特性我们克隆、测序并分析了编码新的IKB蛋白的MAIL基因的表达。

项目成果

Iwasaki, K.: "Isolation, characterization, and cDNA cloning of chicken turpentine-induced protein, a new member of the scavenger receptor cystein-rich (SRCR) family of proteins"J. Biol. Chem.. 276. 9400-9405 (2001)

Iwasaki, K.：“鸡松节油诱导蛋白的分离、表征和 cDNA 克隆，该蛋白是清道夫受体富含半胱氨酸 (SRCR) 蛋白家族的新成员”J.

Kitamura, H.: "MAIL, a novel nuclear IkB protein that potentiates LPS-induced IL-6 production"FEBS Lett.. 485. 53-56 (2000)

Kitamura, H.：“MAIL，一种新型核 IkB 蛋白，可增强 LPS 诱导的 IL-6 产生”FEBS Lett.. 485. 53-56 (2000)

Nawa,A., et al.: "Tumor metastasis-associated human MTA1 gene : its deduced protein sequence, localization, and association with breast cancer cell proliferation using antisense phosphorothioate oligonucleotides"J Cell Biochem.. 79. 202-212 (2000)

Nawa,A., et al.：“肿瘤转移相关的人 MTA1 基因：使用反义硫代磷酸寡核苷酸推导的蛋白质序列、定位以及与乳腺癌细胞增殖的关联”J Cell Biochem.. 79. 202-212 (2000)

Kitamura, H.: "Bacterial lipopolysaccharide induced mRNA expression of a IkB MAIL mediated through Toll-like receptor 4"J. Vet. Med. Sci. (印刷中).

Kitamura, H.：“细菌脂多糖通过 Toll 样受体 4 介导的 IkB MAIL 的 mRNA 表达”J. Vet。

Kitamura,H. ら: "MAIL.a novel nuclear IkB protein that potentiates LPS-induced IL-6 production."FEBS Lett.. 485. 53-56 (2000)

Kitamura, H. 等人：“MAIL.一种新型核 IkB 蛋白，可增强 LPS 诱导的 IL-6 产生。”FEBS Lett.. 485. 53-56 (2000)