Isolation of model mouse generating Ewing sarcoma and analysis of the mechanism of sarcoma-genesis

尤文肉瘤小鼠模型的分离及肉瘤发生机制分析

• 批准号: 13671526
• 负责人: YOSHIDA Koichi
• 金额: $ 2.18万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671526/
• 关键词: sarcoma; mouse model of disease; chromosome translocation; fusion gene; cre-loxP DNA recombination; transgenic mouse; cancer; ETS transcription factor; Cre-1oxP DNA組み換え

The purposes of this study is to isolate a model mouse generating Ewing sarcoma and peripheral neuroectodermal tumor (PNET) by expressing the sarcoma-specific EWS-FLI fusion oncogene in the mouse tissue limited for generation of sarcoma, and to identify the genes regulated by tumor-specific transcription factor, the product of EWS-FLI fusion oncogene. Two EWS-FLI transgenic mouse strains with fusion oncogene were isolated and found to be reproductive and able to grow up without any significant failures. To allow DNA recombination in vivo by the Cre-loxP techniques, EWS-FLI mice were crossbred with the Cre transgenic mice manipulated to express the DNA recombinase in neural crest-derived cells. Since mice with the recombined DNA yielded at lower frequency, we were currently unable to determine whether tumor is generated or not. By breeding a different line of EWS-FLI mouse strains, the birth of recombinant mice will be improved. On the other hand, we found several genes whose expressions were altered with the sarcoma-specific fusion oncogene but not with its normal counterpart. One is the extra-cellular matrix component Tenascin-C associated with tumor progression. The other is transcription factor Id2. Their biological significance in Ewing sarcoma was investigated (Genes Chromosomes & Cancer 2003 ; Oncogene 2002). Also we found up-regulation of c-myc and down-regulation of TGF-beta2 and Smad3 expressions by fusion oncogene. These may be associated with higher proliferating potential of Ewing sarcoma cells. To elucidate the mechanism of sarcoma-genesis, further analysis of target genes will be required.

本研究的目的是通过在限制肉瘤发生的小鼠组织中表达肉瘤特异性EWS-FLI融合癌基因，分离产生Ewing肉瘤和周围神经外胚层肿瘤（PNET）的模型小鼠，并鉴定EWS-FLI融合癌基因产物肿瘤特异性转录因子调控的基因。分离出两株EWS-FLI转基因融合癌基因小鼠，发现它们具有繁殖能力，并且能够在生长过程中没有明显的失败。为了利用Cre- loxp技术在体内进行DNA重组，我们将EWS-FLI小鼠与Cre转基因小鼠杂交，使其在神经冠源性细胞中表达DNA重组酶。由于重组DNA的小鼠产生的频率较低，我们目前无法确定是否会产生肿瘤。通过培育不同的EWS-FLI小鼠品系，将改善重组小鼠的出生。另一方面，我们发现有几个基因的表达随肉瘤特异性融合癌基因而改变，而不随其正常对应基因而改变。一种是与肿瘤进展相关的细胞外基质成分Tenascin-C。另一个是转录因子Id2。研究了它们在尤文氏肉瘤中的生物学意义（基因、染色体和癌症2003；癌基因2002）。我们还发现融合癌基因可上调c-myc，下调tgf - β 2和Smad3的表达。这些可能与尤文氏肉瘤细胞较高的增殖潜能有关。为了阐明肉瘤发生的机制，需要进一步分析靶基因。

项目成果

Nishimori H, Sasaki Y, Yoshida K, Irifune H, Zenbutsu H, Tanaka T, Aoyama T, Hosaka T, Kawaguchi S, Wada T, Ishii S, Hata J, Toguchida J, Nakamura Y, Tokino T: "The Id2 gene is a novel target of transcriptional activation by EWS-ETS fusion proteins in Ewi

Nishimori H、Sasaki Y、Yoshida K、Irifune H、Zenbutsu H、Tanaka T、Aoyama T、Hosaka T、Kawaguchi S、Wada T、Ishii S、Hata J、Toguchida J、Nakamura Y、Tokino T：“Id2 基因是

Yabe H, Fukuma M, Urano F, Yoshida K, Kato S, Toyama Y, Hata J, Umezawa A: "Lack of matrix metalloproteinase (MMP)-l and MMP-3 expression in Ewing sarcoma may be due to loss of accessibility of the MMP regulatory element to the specific fusion protein in

Yabe H、Fukuma M、Urano F、Yoshida K、Kato S、Toyama Y、Hata J、Umezawa A：“尤文肉瘤中基质金属蛋白酶 (MMP)-1 和 MMP-3 表达的缺乏可能是由于缺乏

Hiromitsu Hiroumi: "Expression of E1AF/PEA3, an ETS-related transcription factor in human Non-small-cell lung cancers : its relevance in cell motility and invasion"Int. J. Cancer. 93. 786-791 (2001)

Hiromitsu Hiroumi：“人类非小细胞肺癌中 E1AF/PEA3（一种 ETS 相关转录因子）的表达：其与细胞运动和侵袭的相关性”Int。