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

鉴定与预后不良的癌症的发生和进展有关的新癌基因。

• 批准号: 09670145
• 负责人: UCHIDA Kazuhiko
• 金额: $ 1.92万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670145/
• 关键词: biliary tract cancer; ovarian cancer; neuroblastoma; gastric cancer; DNA microarray; array CGH; prognosis; 1q23 amplification; 難治がん; 浸潤; 転移; マイクロアレイ; 難治癌; 遺伝子; CGH; 胆道癌; 卵巣癌

(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

（1997）尽管临床技术的发展，许多癌症的诊断仍然很差。比较基因组杂交（CGH）技术可用于发现与肿瘤发生有关的新的癌基因和抑癌基因。1)对30例新鲜冰冻胆囊癌组织进行CGH。12p染色体获得与III期和IV期相关（P <0.05）。因此，12 p的增加可能是胆囊癌的一个潜在的预后因素。2)卵巢癌：为了发现耐药肿瘤的基因变化，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的增益以及2p和15q的损失与顺铂耐药表型相关。获得紫杉醇耐药的细胞株在MDR基因所在的7q染色体上有增加。Southern杂交证实MDR基因扩增。目前的研究结果表明，这些染色体 ...更多信息 染色体获得和丢失可能是预测卵巢癌患者在顺铂和/或紫杉醇化疗前耐药的潜在指标。3)神经母细胞瘤：我们对24例神经母细胞瘤进行了CGH和双色FISH，以确定与进行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的新染色体获得。此外，通过使用粘粒克隆的FISH分析，1q21-q25增益被缩小到1q23。提示1q23区DNA扩增可能在进展期神经母细胞瘤的发生发展中起一定作用。为了阐明退行性和进行性神经母细胞瘤的生物学特征，对67例神经母细胞瘤患者进行了CGH。所有消退肿瘤（1 - 3期和4期）的CGH数据显示染色体的整个部分的全染色体畸变。另一方面，进行性肿瘤显示染色体区域部分的染色体获得和丢失。我们的数据表明，神经母细胞瘤分为两个生物学上不同的组，其中一组显示进行性疾病，显示进行性疾病，其中有部分染色体的变化，而另一个模仿先进的3/4期神经母细胞瘤，但显示退行性疾病，其中有整个染色体畸变。（1999）DNA微阵列的最新进展使我们能够对遗传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。我们尝试建立一种高分辨率、高灵敏度、高定量能力的微阵列计算全息图，并将其应用于肿瘤组织中癌基因的定位。我们的阵列CGH显示DNA拷贝数在1至10，000个拷贝范围内的定量分析。我们在每个细胞中检测到10个基因拷贝。我们还使用含有4，000个已知基因的cDNA的微阵列进行了胃癌的基因表达监测，并鉴定了多种在胃癌中特异表达的基因。少

项目成果

Kudoh, K., Takano, M., Yoshida, S., Mano, Y., Yamamoto, K., Ishii, K., Kita, T., Kikuchi, Y., Nagata, I., Miwa, M., and Uchida, K.: "Gains of 1q21-q22 and 13q12-q14 are potential indicators for resistance to cisplatin-based chemotherapy in ovarian cancer

Kudoh，K.，高野，M.，吉田，S.，马野，Y.，山本，K.，石井，K.，北，T.，菊池，Y.，永田，I.，美轮，M.，

J. Fang, S. Kushida, R. Feng, M. Tanaka, H. Kikukawa, T. Kawamura, K. Uchida and M. Miwa.: "Integration of HTLV-1 provirus into mouse transforming growth factor-α gene."Biochem. Biophys. Res. Commun.. 233. 792-795 (1997)

J. Fang、S. Kushida、R. Feng、M. Tanaka、H. Kikukawa、T. Kawamura、K. Uchida 和 M. Miwa.：“HTLV-1 原病毒与小鼠转化生长因子-α 基因的整合。”生物化学研究。233。792-795（1997）

Hanai,S.et al: "Genomic Organization of Drosophila Poly(ADP-ribose)Polymerase and Distribution of Its mRNA during Development" J.Biol.Chem.(in press). (1998)

Hanai,S.et al：“果蝇聚（ADP-核糖）聚合酶的基因组组织及其发育过程中 mRNA 的分布”J.Biol.Chem.（出版中）。

Maeda,N.et al: "Inhibition of human T-cell leukomia virus type I replication by antisenso env" Biochem.Biophys.Res.Commun.243. 109-112 (1998)

Maeda,N.等人：“反义环境对人 T 细胞白血病病毒 I 型复制的抑制”Biochem.Biophys.Res.Commun.243。

内田和彦 他: "DNAマイクロアレイを用いたゲノムワイドの遺伝子増幅、欠失の解析"遺伝子医学. 4. 113-117 (1999)

Kazuhiko Uchida 等人：“使用 DNA 微阵列进行全基因组基因扩增和缺失分析”遗传医学。 4. 113-117 (1999)