Chromosome 22q genetic studies in the colon

结肠染色体 22q 遗传学研究

• 批准号: 7590697
• 负责人: Anil K Rustgi
• 金额: $ 12.68万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2008-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7590697
• 关键词: 17p; 18q; 22q; 22q13; Adenomatous Polyposis Coli; Adenomatous Polyps; Allelic Imbalance; Animal Model; Bacterial Artificial Chromosomes; Bacteriophage P1; Behavior; Biochemical; Biological; Biological Assay; Breast; Candidate Disease Gene; Cell Line; Cell physiology; Cessation of life; Chromosomes; Chromosomes, Artificial, Yeast; Clinical; Cloning; Colon; Colon Carcinoma; Colonic Neoplasms; Colorectal; Colorectal Cancer; Complement; Cosmids; Custom; DNA Markers; DNA Sequence; Data; Databases; Development; Disease; Embryonic Development; Epigenetic Process; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Expressed Sequence Tags; Gene Expression; Gene Mutation; Genes; Genetic; Genomics; Genus Cola; Homeostasis; Human; Human Genome Project; Immunohistochemistry; In Situ Hybridization; In Vitro; Individual; Intestines; Length; Link; Loss of Heterozygosity; Maintenance; Malignant - descriptor; Malignant Neoplasms; Messenger RNA; Methylation; Microsatellite Repeats; Modeling; Molecular; Mutation; Nucleotides; Oral; Ovarian; Pathogenesis; Pattern; Physical Chromosome Mapping; Polymerase Chain Reaction; Premalignant; Property; Proteins; Relative (related person); Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; SSCP Analysis; Single-Stranded Conformational Polymorphism; Small Interfering RNA; Source; TP53 gene; Technology; Tissues; Tumor Suppressor Genes; United States; Variant; Villus; Work; Xenograft Model; base; cDNA Arrays; cancer genetics; carcinogenesis; chromosome 5q loss; colon cancer cell line; innovation; insight; mRNA Expression; malignant state; novel; outcome forecast; positional cloning; programs; tumor

There are approximately 140,000 new cases of colorectal cancer annually in the United States, with nearly 60,000 deaths related to complications of disease. The model of colorectal cancer development and progression, espoused by Volgelstein and colleagues, represents a paradigm in which to understand cancer genetics. The identification of novel genes involved in proliferation and malignant transformation has been greatly enhanced through cloning after localization of chromosomal regions that are deleted (designated as loss of heterozygosity-LOH- or allelic deletion) during malignant transformation. The colon is no exception and indeed, identification of critical genes such as APC for adenomatous polyposis coli (chromosome 5q), p53 (chromosome 17p), and Smad4 (chromosome 18q). Recognizing that other chromosomal regions are frequently deleted in the progression premalignant to malignant states in the colon, we have meticulously identified a new target region of allelic loss on chromosome 22q that is involved in human colorectal carcinogenesis. Fine genetic and physical mapping with microsatellite DNA markers demonstrates that the interval on chromosome 22q13.31 is less than 1 MB and contains 14 genes (including ESTs). We hypothesize that a tumor suppressor gene(s) exist (s) whose encoded proteins harbor critical genetic, biological and biochemical properties that ultimately is important in the maintenance of intestinal epithelial cell homeostasis and linked to progression to cancer. Therefore, to achieve the identification of the gene through well established technologies and attain its subsequent molecular characterization, we will pursue well-integrated Specific Aims: (1) To determine the mRNA expression of the 14 genes in colon tumors relative to normal colon using a custom made cDNA array; and to narrow the region o fLOH using PCR-SNP technology; (2) To identify the chromosome 22q13 candidate gene(s) from among other genes in the region, each candidate gene will be studied for tumor-specific alterations (mutations) using PCR- SSCP followed by DNA sequencing of SSCP variants, and to investigate for the possibility of epigenetic inactivation employing PCR-methylation assays; and (3) To evaluate gene expression at the mRNA and protein levels during development, states of differentiation and proliferation, and assess correlations with clinical parameters; to evaluate the functional properties of the tumor suppressor gene in vitro and in animal models. These innovative and cohesive studies will permit important mechanistic insights into molecular pathogenesis and create a platform for translational applications.

美国每年大约有140,000例新的结直肠癌病例，近60,000例 与疾病并发症有关的死亡。结直肠癌的发展和进展模型，由 Volgelstein及其同事代表了一个了解癌症遗传学的范式。新颖的识别 通过克隆，与增殖和恶性转化有关的基因已大大增强 被删除的染色体区域的定位（被指定为杂合性 - LOH-或等位基因缺失的损失） 在恶性转化期间。结肠也不例外，实际上，鉴定关键基因（例如APC） 腺瘤性息肉病大肠杆菌（5q染色体），p53（17p染色体）和SMAD4（染色体18Q）。认可 其他染色体区域经常在结肠前对恶性状态的进展中被删除， 我们精心识别了与人类有关的22q染色体上等位基因损失的新目标区域 结直肠癌发生。用微卫星DNA标记物的细遗传和物理映射表明 染色体22q13.31的间隔小于1 Mb，包含14个基因（包括EST）。我们假设一个 肿瘤抑制基因（S）的编码蛋白具有关键遗传，生物学和生化特性 最终，这对于维持肠上皮细胞体内平衡很重要，并且与发展有关 癌症。因此，通过建立良好的技术来实现基因的识别并实现其 随后的分子表征，我们将追求良好的特定目的：（1）确定mRNA 使用定制的cDNA阵列相对于正常结肠的14个基因的表达；并缩小 使用PCR-SNP技术的floh区域； （2）从中间识别染色体22q13候选基因 该地区的其他基因，将使用PCR-将研究每个候选基因的肿瘤特异性改变（突变） SSCP随后进行SSCP变体的DNA测序，并研究表观遗传失活的可能性 采用PCR-甲基化测定； （3）评估在mRNA和蛋白质水平上的基因表达 发展，分化和增殖状态，并评估与临床参数的相关性；评估 在体外和动物模型中，肿瘤抑制基因的功能特性。这些创新和凝聚力的研究 将允许对分子发病机理的重要机理见解，并为翻译创建平台 申请。