Development of Systems for Comprehensive Genetic Diagnosis of Human Cancers and for Early Detection of Cancer-patients.

人类癌症综合基因诊断和癌症患者早期检测系统的开发。

• 批准号: 10470040
• 负责人: SHIMIZU Kenji
• 金额: $ 3.65万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470040/
• 关键词: p107 gene; HD-PTP ; Tyrosine-phosphatase gene; Chromosome 3p21; Tumor suppressor genes; ING1 gene; loss of heterozygosity (LOH); Chromosome 13q34; Genetic diagnosis with peripheral blood; ヘテロ接合性消失; 融合遺伝子特異的分子診断; 活性化変異特異的遺伝子診断; 遺伝子内欠失; チロシンホスファ

We have found a novel tumor-specific mutation in the gene encoding E2F4 transcription regulator in a subset of genetically unstable (MI+) colorectal cancers in 1996. The mutation was found to be a result of the inactivating frame-shift mutation of the hMSH3 gene, a DNA mismatch-repair gene. Thus, the E2F4 gene was identified, for the first time, as a target of the defective mismatch repair function involving hMSH3 protein. This notion has been confirmed bv many supporting papers. Relevant to this result, we have found an interstitial deletion within the human p107 gene, a close relative of the tumor suppressor gene RB, in a diffuse-large B cell lymphoma cell line. We found the deletion was raised by a recombination between two Alu-repeats encompassing about 15 kbp. By our genomic scanning method, we found a novel region of loss of heterozygosity (LOH) at chromosome 14q24-31 in about 30% of renal cell carcinomas and identified two candidate tumor suppressor genes (TSG) in this narrow re … More gion. By conventional microsatellite analysis, we found a high incidence of LOH at chromosome 13q34 in human head and neck cancers. The known candidate TSG, ING1, resides at this site. We clarified the whole genomic structure of the ING1 gene and found 3 inactivating point mutations in 3 primary head and neck cancers with 13q34 LOH.These are the first evidence indicatins that the p107 and ING1 genes function as TSG.Further, we discovered a novel protein tyrosine-phosphatase gene, HD-PTP, as a novel TSG candidate. The gene resides at chromosome 3p21.3, the region thought to contain multiple TSG for many types of human cancer. By similar approaches we found at least 10 TSG candidates at 8 chromosomal regions in 7 cancer/tumor groups. In addition to TSG, we isolated a cDNA encoding a novel POZ-Zn-Finger protein, whose expression led the recipient cells to malignant phenotype, suggesting that the gene is a novel oncogene. The gene locates on 3q26-27 and related to the known oncogene, Bcl-6.As to the molecular diagnosis of cancer, we reported a systematic method for detecting tumor-specific, fused-gene mRNAs in several bone/soft-tissue sarcomas. We have developed a new strategy to quantitate the amounts of free DNA in sera/plasma with a very high sensitivity (about ten thousand-fold higher than conventional methods). Cancer-specific genetic alterations, like as K-ras and p53 mutations, could be detected in these serum-DNA.Thus, promising strategies for early detection of potential cancer-patients were developed with free DNA in plasma/serum. Less

1996 年，我们在一组遗传不稳定 (MI+) 结直肠癌的编码 E2F4 转录调节因子的基因中发现了一种新的肿瘤特异性突变。该突变被发现是 hMSH3 基因（一种 DNA 错配修复基因）失活移码突变的结果。因此，E2F4基因首次被鉴定为涉及hMSH3蛋白的缺陷错配修复功能的靶标。这一观点已被许多支持论文所证实。与这一结果相关的是，我们在弥漫性大 B 细胞淋巴瘤细胞系中发现了人类 p107 基因（肿瘤抑制基因 RB 的近亲）内的间质缺失。我们发现缺失是由两个大约 15 kbp 的 Alu 重复序列之间的重组引起的。通过我们的基因组扫描方法，我们在约 30% 的肾细胞癌中发现了染色体 14q24-31 上的一个新的杂合性丢失 (LOH) 区域，并在这个狭窄的区域中鉴定了两个候选肿瘤抑制基因 (TSG)。通过传统的微卫星分析，我们发现人类头颈癌中染色体13q34处的LOH发生率很高。已知的候选 TSG ING1 位于此站点。我们阐明了 ING1 基因的全基因组结构，并在 3 个原发性头颈癌 13q34 LOH 中发现了 3 个失活点突变。这是第一个证据表明 p107 和 ING1 基因具有 TSG 功能。此外，我们发现了一个新的蛋白酪氨酸磷酸酶基因 HD-PTP，作为新的 TSG 候选基因。该基因位于染色体 3p21.3，该区域被认为含有多种 TSG，可导致多种类型的人类癌症。通过类似的方法，我们在 7 个癌症/肿瘤组的 8 个染色体区域发现了至少 10 个 TSG 候选者。除了 TSG 之外，我们还分离了编码新型 POZ-Zn-Finger 蛋白的 cDNA，其表达导致受体细胞出现恶性表型，表明该基因是一种新型癌基因。该基因位于3q26-27，与已知的癌基因Bcl-6相关。在癌症的分子诊断方面，我们报道了一种系统方法，用于检测几种骨/软组织肉瘤中肿瘤特异性融合基因mRNA。我们开发了一种新策略，以非常高的灵敏度（比传统方法高出约一万倍）对血清/血浆中游离 DNA 的量进行定量。癌症特异性基因改变，如 K-ras 和 p53 突变，可以在这些血清 DNA 中检测到。因此，利用血浆/血清中的游离 DNA 开发了早期检测潜在癌症患者的有前景的策略。较少的

项目成果

Horibe, K., et al: "Prognostic Factors in Childhood Acute Lymphoblastic Leukemia in Japan."Inter.J.Hematol.. 72. 61-68 (2000)

Horibe, K., 等人：“日本儿童急性淋巴细胞白血病的预后因素。”Inter.J.Hematol.. 72. 61-68 (2000)

Shibuya Kazuko: "Physical and functional association of αLβ2 integrin(LFA-1) with DNAM-1 adhesion molecule"Immunity. 11. 1-20 (1999)

涩谷和子：“αLβ2 整合素 (LFA-1) 与 DNAM-1 粘附分子的物理和功能关联”免疫。 11. 1-20 (1999)

Nishiyama,M.: "11p15 translocations involving the NUP98 gene in childhood therapy-related acutemyeloid leukemia/myelodysplastic syndrome."Genes Chromoso.Cancer. 26. 215-220 (1999)

Nishiyama, M.：“儿童治疗相关的急性髓性白血病/骨髓增生异常综合征中涉及 NUP98 基因的 11p15 易位。”基因 Chromoso.癌症。

Horibe,K.: "Prognostic Factors in Childhood Acute Lymphoblastic Leukemia in Japan."Intern.J.Hematol.. 72. 61-68 (2000)

Horibe,K.：“日本儿童急性淋巴细胞白血病的预后因素。”Intern.J.Hematol.. 72. 61-68 (2000)

Yabe, Y., et al: "Human papillomavirus-5b DNA integrated in a metastatic tumor : cloning, nucleotide sequence and genomic organization."Int.J.Cancer. 80. 334-335 (1999)

Yabe, Y., 等人：“整合到转移性肿瘤中的人乳头瘤病毒 5b DNA：克隆、核苷酸序列和基因组组织。”Int.J.Cancer。