Identification of novel oncogenes invloved in onset and progression of cancers with poor prognosis.

尽管临床技术有所发展，但许多癌症的预后仍然很差。比较基因组杂交技术（CGH）可用于鉴定与肿瘤发生有关的新型癌基因和抑癌基因。1)胆道癌；对30例新鲜冷冻胆囊癌组织进行CGH。染色体增加12p与III期和IV期相关（P<0.05）。因此，12p的增加可能是胆囊癌的一个潜在预后因素。2)卵巢癌；为了发现耐药肿瘤的遗传变化，对21例原发性卵巢癌和部分细胞系进行了CGH。我们发现染色体区域1p、1q和19p的增加，以及2p和15q的减少与顺铂耐药表型有关。获得紫杉醇抗性的细胞系在MDR基因所在的染色体上增加了7q。经southern blot分析证实MDR基因扩增。目前的研究结果表明，这些染色体的增加和减少可能是预测卵巢癌患者在顺铂和/或紫杉醇化疗前耐药性的潜在指标。3)神经母细胞瘤；我们对24个神经母细胞瘤进行了CGH和双色FISH，以确定与进展性神经母细胞瘤相关的遗传畸变。在所有进行性4期神经母细胞瘤中发现了1q21-q25的新染色体增加。此外，通过使用cosmid克隆进行FISH分析，将1q21-q25的增益缩小到1q23。这些结果提示1q23的DNA扩增可能在晚期进行性神经母细胞瘤的发展中起作用。（1998）为了明确退行性和进行性神经母细胞瘤的生物学特征，我们对67例神经母细胞瘤患者进行了CGH。所有退行性肿瘤（e1-3期和4s期）的CGH数据均显示整部分染色体畸变。另一方面，进展性肿瘤在染色体的局部部分表现出染色体的获得和损失。我们的数据表明，神经母细胞瘤在生物学上分为两种不同的组，其中一种表现为进行性疾病，表现为部分染色体改变的进行性疾病，而另一种模拟晚期3/4期神经母细胞瘤，但表现为退行性疾病，表现为整个染色体畸变。（1999） DNA微阵列的最新进展使我们能够对基因改变进行全基因组分析，包括DNA拷贝数变化、突变和癌症中的基因表达。我们试图建立基于阵列的高分辨率、高定量能力和灵敏度的CGH，并将这种新的强大方法应用于癌症中推定的致癌基因的定位。我们的阵列CGH显示了DNA拷贝数的定量分析，范围从1到10,000拷贝。我们在每个细胞中检测到10个基因拷贝。我们还利用含有4000个已知基因cdna的微阵列对胃癌进行了基因表达监测，发现了许多在胃癌中特异性表达的基因。少

(1997) In spite of the development of clinical techniques, the prognoses of many cancers are still poor. Comparative genomic hybridization (CGH) is useful for identification of novel oncogenes and tumor suppressor genes involved in the carcinogenesis. 1) Biliary tract cancer ; CGH was performed on 30 fresh frozen tissues of gallbladder cancers. Chromosomal gain of 12p was associated with stage III and IV (P<0.05). Therefore, gain of 12p may be a potential prognostic factor of gallbladder cancers. 2) Ovarian cancer ; In order to find genetic changes in drug-resistant tumors, CGH was performed on 21 primary ovarian cancers and some cell lines. We found gains in chromosomal regions 1p, 1q and 19p, and losses in 2p and 15q to be related to the cisplatin-resistant phenotype. The cell lines which acquired the taxol-resistance had chromosomal gain of 7q where MDR gene was located. The amplification of MDR gene was confirmed by southern blot analysis. Present findings suggest that these chromo … More somal gains and losses may be potential indicators for prediction of resistance in ovarian cancer patients before cisplatin- and/or taxol-based chemotherapy. 3) Neuroblastoma ; We performed CGH on 24 neuroblastomas and dual-color FISH to identify genetic aberrations associated with progressive neuroblastoma. A novel chromosomal gain at 1q21-q25 was found in all of progressive stage 4 neuroblastomas. Furthermore, by FISH analysis using cosmid clones, the 1q21-q25 gain was narrowed to 1q23. These results suggest that DNA amplification at 1q23 may play a role in the development of progressive neuroblastoma in advanced stage.(1998) To clarify the biological characteristics of regressive and progressive neuroblastomas, CGH was performed on 67 patients with neuroblastomas. The CGH data of all regressive tumors (stage1-3 and 4s) revealed whole-chromosome aberrations of whole portion of chromosome. On the other hand, progressive tumors revealed chromosomal gains and losses on regional portion of chromosome. Our data suggest that neuroblastomas are classified into two biologically different groups, one of which displays progressive disease which displays progressive disease which has partial chromosomal changes, whereas the other mimics advanced stage 3/4 neuroblastoma but displays regressive disease which has whole chromosomal aberrations.(1999) Recent advances in DNA microarray allow us genome-wide analysis of genetic alterations including DNA copy number changes, mutations, and gene expression in cancers. We tried to establish array-based CGH with high resolutions, high quantitative capability and sensitivity, and applied this novel powerful method to mapping the putative oncogenes in cancers. Our arrayed CGH showed quantitative analysis of DNA copy number in the range from 1 to 10,000 copies. We detected 10 copies of the gene per cell. We also performed gene expression monitoring of gastric cancer using microarray with 4,000 cDNAs of known genes and identified many kinds of genes that were specifically expressed in gastric cancer. Less