Rapid quantification system of DNA copy number for the specific molecular targeting area in stomach cancer tissue using by quantitative real time PCR assay.

通过实时定量 PCR 测定，对胃癌组织中特定分子靶向区域的 DNA 拷贝数进行快速定量系统。

• 批准号: 16591362
• 负责人: SUZUKI Seiji
• 金额: $ 1.79万
• 依托单位: Nippon Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591362/
• 关键词: Quantitative microsatellite analysis; Cancer chemotherapy; Active immunotherapy; ZNF217; AIB-1; c-MYC; Highly sensitive in situ hibridization; MAGE-A10; Cancer testis antigen; MAGE-A family; TRAG-3; Monoclonal antibody; in situ hybridization /

In 2004, our group almost collected the information about the gene specific areas which were associated with the molecular targeting therapy, active immunotherapy and cancer chemotherapy. The quantification of ZNF217 DNA copy number which had been succeeded in previously was analyzed for more clinical samples. In 2005, using by quantitative real time microsatellite analysis (QuMA), the DNA copy number of AIB-1 in chromosome 20q and c-Myc in chromosome 8q were observed in stomach cancer. Further, our group focused on the MAGE-A family (MAGE-A10・MAGE-A3) as cancer testis antigens (CTA) and TRAG-3 related to the drug resistance of Taxan and the specific target for the active immunotherapy. Immunohistochemically, by using monoclonal antibody 6C1, MAGE-A family protein expression in cases with stomach cancer metastasized to the liver were significantly observed (about 70%) than those with early stage cancer and those with advanced stage cancer who were no recurrent for three years after surgery. In 2006, our group detected the MAGE -A10 mRNA signals by highly sensitive in situ hybridization (ISH) using cRNA probes since no specific monoclonal antibody for MAGE-A10 were not available. MAGE-A family expression by 6C1 and MAGE-A10 mRNA expression by highly sensitive ISH were well concordant. We think that IHC and highly sensitive ISH using cRNA probes were useful for the screening for the selection of the gene specific areas which could be measured DNA copy number by QuMA.

2004年，本课题组收集了与分子靶向治疗、主动免疫治疗和肿瘤化疗相关的基因特异性区域的信息。对ZNF 217 DNA拷贝数的定量方法进行了改进，并对更多的临床样本进行了分析。2005年，应用微卫星真实的时间定量分析（QuMA）技术，观察了胃癌组织中20号染色体上AIB-1和8号染色体上c-Myc基因的DNA拷贝数。此外，我们的小组集中在MAGE-A家族（MAGE-A10·MAGE-A3）作为癌症睾丸抗原（CTA）和TRAG-3与紫杉醇耐药性和主动免疫治疗的特异性靶点。免疫组化结果显示，MAGE-A家族蛋白在胃癌肝转移患者中的表达率为70%，明显高于早期胃癌和术后3年无复发的晚期胃癌。2006年，由于没有针对MAGE-A10的特异性单克隆抗体，我们小组使用cRNA探针通过高灵敏度原位杂交（ISH）检测了法师-A10 mRNA信号。通过6C 1的MAGE-A家族表达和通过高灵敏度ISH的MAGE-A10 mRNA表达具有良好的一致性。我们认为，使用cRNA探针的IHC和高灵敏度ISH可用于筛选可通过QuMA测量DNA拷贝数的基因特异性区域。

项目成果

Comparative study between DNA copy number abberations determined by quantitative real time microsatellite analysis and clinical outcome in patients with stomach cancer

实时定量微卫星分析确定的DNA拷贝数畸变与胃癌患者临床结果的比较研究

• 发表时间: 2004
• 期刊: Clinical Cancer Research 10
• 作者: Suzuki S;Egami K;Sasajima K;Mohammad G;Shimizu H;Watanabe H;Hasegawa H;Iida S;Matsuda T;Okihama Y;Hosone M;Shimizu K;Kawanami O;Tajiri T
• 通讯作者: Tajiri T

Comparative study between DNA copy number aberrations determined by quantitative microsatellite analysis and clinical outcome in patients with stomach cancer

定量微卫星分析确定的DNA拷贝数畸变与胃癌患者临床结果的比较研究

• 发表时间: 2004
• 期刊: Clinical Cancer Rpsearch 10
• 作者: Suzuki S;Egami K;Sasajima K;Mohammad G;Shimizu H;Watanabe H;Hasegawa H;Iida S;Matsuda T;Okihama Y;Hosone M;Shimizu K;Kawanami O;Tajiri T
• 通讯作者: Tajiri T