SOMATIC GENETIC ALTERATIONS IN GASTROINTESTINAL CANCER

胃肠癌的体细胞遗传改变

• 批准号: 2089589
• 负责人: MANUEL PERUCHO
• 金额: $ 19.61万
• 依托单位: CALIFORNIA INSTITUTE OF BIOLOGICAL RES
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-30 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2089589
• 关键词: CHO cells; adenocarcinoma; adenoma; aneuploidy; colorectal neoplasms; genetic mapping; genome; human genetic material tag; human subject; hybrid cells; metastasis; molecular oncology; neoplasm /cancer diagnosis; neoplastic process; neoplastic transformation; nucleic acid sequence; oncogenes; polymerase chain reaction; restriction fragment length polymorphism; tumor suppressor genes

We have adapted a DNA fingerprinting approach to detect tumor specific genomic differences. The approach is based in the comparison of the genome fingerprints of normal and tumor tissues from the same individuals, after the amplification of DNA sequences by the Arbitrary Priming Polymerase Chain Reaction (AP-PCR). The method is able to detect in a single and simple experiment, both quantitative and qualitative genomic differences associated with neoplastic transformation. It also allows the cloning in a single step of these altered DNA sequences, including those deleted in the tumor cells. We propose to use this approach as a molecular alternative to the cytogenetic analysis of solid tumors; to estimate the extent and compare the spectrum of genetic alterations occurring in diverse tumors of the digestive tract at different stages of tumor progression; to explore its potential value as prognostic indicator for Cancer of the colon and rectum; and to search for unknown cancer genes in colorectal and other gastrointestinal tumors. We will test the hypothesis that the relative values of genetic damage obtained by the AP-PCR method reflect the degree of aneuploidy of the cancer cells, and that this information might have value for cancer diagnosis and prognosis. The existence of uncharacterized cancer genes involved in the development and/or progression of defined gastrointestinal neoplasms will be searched by isolating and characterizing the chromosomal localization of DNA sequences representing recurrent genetic alterations in these tumors. Genetic alterations will be identified in a panel of about 200 tumors of the gastrointestinal tract including adenomas and carcinomas of the colon and rectum, pancreas and stomach, by comparison of the AP-PCR patterns of tumor versus normal tissues. Genomic alterations specific to the metastatic process of any of these tumors will be searched by comparing primary versus metastatic tumors by the same approach (specific aim 1). We will continue the follow-up of a subset of these tumors (about 100 colorectal carcinomas), which have been already followed for an average of over 3 years for recurrence and survival. These tumors have been already characterized for mutations in the ras oncogene and in the p53 tumor suppressor gene. The relative values of genetic damage obtained for these colorectal carcinomas by the AP-PCR method will be correlated with ras and p53 gene mutations and recurrence and survival rates (specific aim 2). The chromosomal localization of DNA fragments corresponding to recurrent tumor specific genetic alterations will be determined by AP-PCR amplification of panels of human/hamster cell hybrids. When appropriate, these DNA fragments will be cloned and sequenced to further characterize by standard PCR approaches their chromosomal localization and their presence and frequency in other tumors (specific aim 3).

我们采用了DNA指纹法来检测肿瘤特异性 基因组差异这种方法是基于基因组的比较 来自同一个体的正常和肿瘤组织的指纹， 用任意引物聚合酶扩增DNA序列 链式反应（AP-PCR）。该方法能够在单个和 简单实验，定量和定性基因组差异 与肿瘤转化有关。它还允许克隆 这些改变的DNA序列的一个步骤，包括那些删除的， 肿瘤细胞。 我们建议使用这种方法作为分子替代的 实体瘤的细胞遗传学分析;估计范围并比较 发生在不同肿瘤中的遗传改变谱 消化道肿瘤进展的不同阶段;探讨其 作为结肠癌预后指标的潜在价值， 直肠;并寻找未知的癌症基因在结肠直肠和其他 胃肠道肿瘤 我们将检验遗传损伤的相对值 通过AP-PCR方法获得的非整倍体的大小反映了非整倍体的程度。 这些信息可能对癌症有价值 诊断和预后。存在不确定的癌症基因 参与定义的胃肠道疾病的发展和/或进展 肿瘤将通过分离和表征染色体 代表复发性遗传改变的DNA序列的定位 在这些肿瘤中。 基因改变将在一组约200个肿瘤中被鉴定， 胃肠道，包括结肠腺瘤和结肠癌 和直肠，胰腺和胃，通过比较的AP-PCR模式， 肿瘤与正常组织的对比。基因组改变特异于 任何这些肿瘤的转移过程将通过比较 原发性肿瘤与转移性肿瘤通过相同的方法（具体目标1）。 我们将继续对这些肿瘤的一个子集（约100例）进行随访。 结直肠癌），已经平均随访了 3年以上复发和生存。这些肿瘤已经 以ras癌基因突变和p53肿瘤为特征 抑制基因获得的这些遗传损伤的相对值 通过AP-PCR方法检测结直肠癌与ras相关， p53基因突变与复发率和生存率的关系（具体目标2）。 复发性大肠癌DNA片段的染色体定位 将通过AP-PCR确定肿瘤特异性遗传改变 扩增人/仓鼠细胞杂交体组。在适当的时候， 这些DNA片段将被克隆和测序，以进一步表征 通过标准的PCR方法，它们的染色体定位和它们的 在其他肿瘤中的存在和频率（具体目标3）。