Study on the induction mechanism of exon skipping and tumorigenesis by 12-repeats

12次重复诱导外显子跳跃和肿瘤发生机制的研究

• 批准号: 09460135
• 负责人: ISHIGURO Naotaka
• 金额: $ 8.06万
• 依托单位: Obihiro University of Agriculture and Veterinary Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460135/
• 关键词: c-myb; lymphoma; tumour; exon; repeat; insertion; DNA; RT-PCR

We found a tandemly repeated 12-nucleotide motif (TGATCTGCCCGT) inserted into the c-myb gene exon 9 in a bovine T-lymphoma cell line BTL-26 which has been established from a calf type of bovine sporadic lymphosarcomas. We investigated the presence of 12-repeat motifs in the other T-lymphoma cell lines from bovine sporadic lymphosarcomas, induction mechanisms of exon 9 skipping in c-myb gene by the 12-repeat motifs and any relationship of turnorigenesis of bovine T-lymphoma and high-transcription-activating activity of c-Myb protein.1. Insertion of a 12-nucleotide motif was found in the c-myb gene exon 9 from three T-lymphoma cell lines (pr1818 and Pr2410) including BTL-26. In addition to 12-repeat, the different 9-nucletide motifs (TCTGGGCGT) was also observed in each cell line. The number of retitive unit ranged from 4 to 17 copies.2. To know whether deficiency in DNA loop repair is involved in the instability of the repeat, abilities to bind and correct the loop structure in nuclear extracts were examined. The data suggest that instability of the 12-repeat in bovine T-lymphoma cells might be independent of deficiency of DNA loop repair function.3. The 12-repeat contained the binding sites (CAACG) for c-Myb and had the ability to transactivate reporter gene in the cells expressing c-Myb or v-Myb.4. It appeared that the deletion of c-myb gene exon 9 was induced by the insertion of the the 12-repeat rather than by the interaction between 12-repeat and c-Myb from the mini-gene plasmid analyses.5. The close relationship between the c-Myb exon 9 skipping and tumorigensesis of bovine T-lymphoma remains unsolved in this study.

我们在牛T淋巴瘤细胞系BTL-26中发现了一个插入c-myb基因外显子9的重复序列(TGATCTGCCCGT)，该细胞系是从小牛型散发性淋巴肉瘤建立的。我们研究了牛散发性淋巴肉瘤的其他T淋巴瘤细胞系中12重复基序的存在，12重复基序诱导c-myb基因外显子9跳跃的机制，以及牛T淋巴瘤的转归与c-Myb蛋白高转录激活活性的关系。在包括BTL-26在内的三个T淋巴瘤细胞系(Pr1818和Pr2410)的c-myb基因外显子9中发现了12个核苷酸基序的插入。除了12个重复序列外，在每个细胞系中还观察到不同的9-核苷酸基序(TCTGGGCGT)。退役单位数量从4份到17份不等。为了了解DNA环修复缺陷是否与重复序列的不稳定性有关，我们检测了核提取液中结合和纠正环结构的能力。这些数据表明，牛T淋巴瘤细胞中12个重复序列的不稳定性可能与DNA环修复功能的缺陷无关。该12重复序列含有c-Myb结合位点(CAACG)，在表达c-Myb或v-Myb的细胞中具有反式激活报告基因的能力。结果表明，c-myb基因外显子9的缺失可能是由12个重复序列的插入引起的，而不是由c-Myb与12个重复序列的相互作用引起的。在本研究中，c-Myb外显子9跳过与牛T淋巴瘤发生之间的密切关系仍未得到解决。

项目成果

Toshie Shinagawa: "Instability of the 12-nucleotide repeat in c-myb gene of bovine T-lympoma cells"The Journal of Veterinary Medical Science. 59. 1071-1074 (1998)

Toshie Shinakawa：“牛 T 淋巴瘤细胞 c-myb 基因中 12 核苷酸重复的不稳定性”《兽医医学科学杂志》。

Yideyuki Onodera: "Characterization of differentially expressed gene in the bovine T lymphoma cell line"Veterinary Immunology and Immunopathology. 62. 209-219 (1998)

Yideyuki Onodera：“牛 T 淋巴瘤细胞系差异表达基因的表征”兽医免疫学和免疫病理学。

Toshie Shinagawa: "Instability of the 12-nucleotide repeat in c-myb gene of bovine T-lymphoma cells" The Journal of Veterinary Medical Science. 59・11. 1071-1074 (1997)

Toshie Shinakawa：“牛T淋巴瘤细胞c-myb基因中12个核苷酸重复的不稳定性”兽医医学杂志59・11（1997）。

Toshie Shinagawa: "Instability of the 12-nucleotide repeat in c-myb gene of bovine T-lymphoma cells"The Journal of Veterinary Medical Science. 59・11. 1071-1074 (1998)

Toshie Shinakawa：“牛T淋巴瘤细胞c-myb基因中12个核苷酸重复的不稳定性”兽医医学杂志59・11（1998）。

Hideyuki Onodera: "Characterization of defferentially expressed gene in the bovine T lymphoma cell line"Veterinary Immunology and Immunopathology. 62. 209-219 (1998)

Hideyuki Onodera：“牛 T 淋巴瘤细胞系中差异表达基因的特征”兽医免疫学和免疫病理学。