New therapeutic strategy using specific fusion genes of bone and soft-tissue sarcomas

利用骨和软组织肉瘤特异性融合基因的新治疗策略

• 批准号: 11671432
• 负责人: YOSHIKAWA Hideki
• 金额: $ 2.24万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671432/
• 关键词: Synovial sarcoma; fusion gene; chemotherapy; lung metastasis; tumor; 骨軟部肉腫; ユーイング肉腫; 染色体; アンチセンス; RT-PCR; 予後

1. We developed two polyclonal antibodies against the N-terminal part and for the C-terminal part of the SYT-SSX2 protein. These antibodies also detected the native protein of 60 kDa in the lysate of a human synovial sarcoma cell line (HS-SY41) with immunoprecipitation, and in extracts of human synovial sarcomas with western blot analysis2. The molecular weight of the SYT-SSX fusion gene product was 60 kDa in human synovial sarcoma3. The NIH3T3 cells transfected with SYT-SSX2 showed a morphological change and colony-forming activity. Both NIH3T3 cells transfected with SYT-SSX1 and 2 produced tumors in nude mice, when the 5xl0^5 cells were inoculated into nude mice4. We developed the antisense oligo DNAs against SYT-SSX fusion gene, which inhibited proliferation of synovial sarcoma cells, but not fibrosarcoma cells5. In a 22-year-old pregnant woman with synovial sarcoma, the SYT-SSX fusion gene transcript was detected by nested polymerase chain reaction (PCR) in the peripheral blood collected before biopsy. The SYT-SSX fusion gene transcript was not detected after chemotherapy and from the peripheral blood from two osteosarcoma patients. These findings suggest that circulating tumor cells should be monitored because they may serve as a prognostic indicator for synovial sarcoma

1.我们开发了两个多克隆抗体的N-末端部分和SYT-SSX 2蛋白的C-末端部分。这些抗体还用免疫沉淀法在人滑膜肉瘤细胞系（HS-SY 41）的裂解物中检测到60 kDa的天然蛋白，并用蛋白质印迹分析2在人滑膜肉瘤的提取物中检测到60 kDa的天然蛋白。SYT-SSX融合基因产物在人滑膜肉瘤中的分子量为60 kDa 3。转染SYT-SSX 2的NIH 3 T3细胞表现出形态学改变和集落形成活性。当将5x10^5个细胞接种到裸鼠中时，用SYT-SSX 1和2转染的NIH 3 T3细胞在裸鼠中均产生肿瘤4。我们开发了针对SYT-SSX融合基因的反义寡核苷酸，其抑制滑膜肉瘤细胞的增殖，但不抑制纤维肉瘤细胞5。在一个22岁的孕妇与滑膜肉瘤，SYT-SSX融合基因转录本检测巢式聚合酶链反应（PCR）在外周血活检前收集。两例骨肉瘤患者化疗后及外周血中均未检测到SYT-SSX融合基因转录本。这些发现提示循环肿瘤细胞应该被监测，因为它们可能作为滑膜肉瘤的预后指标

项目成果

H.Ono: "Expression of smooth muscle calponin in synovial sarcoma"Sarcoma. 3. 107-113 (1999)

H.Ono：“滑膜肉瘤中平滑肌钙调蛋白的表达”肉瘤。

Naka, N.: "Expression of SSX genes in human osteosarcomas"International Journal of Cancer. 98. 640-642 (2002)

Naka, N.：“SSX 基因在人骨肉瘤中的表达”国际癌症杂志。

I.Kudawara: "Giant cell tumor arising from the spinous process of the sacrum"Eur Radiol. 10・7. 1202 (2000)

I.Kudawara：“骶骨棘突产生的巨细胞瘤”Eur Ra​​diol 10・7（2000）。

T.Tomita: "Transcription factor decoy for NFkappa B inhibits cytokine and adhesion molecule expressions in synovial cells derived from rheumatoid arthritis"Rheumatology. 39・7. 749-757 (2000)

T. Tomita：“NFkappa B 的转录因子诱饵抑制类风湿性关节炎滑膜细胞中的细胞因子和粘附分子表达”《风湿病学》39·7（2000）。

名井 陽: "骨肉種の遺伝子異常"関節外科. 21. 598-601 (2002)

Yo Nai：“骨肉瘤的遗传异常”关节外科。21. 598-601 (2002)