A novel mutation in the T8 microsatellite of 3-UTR CDK2-AP1 gene in MSI CRC

MSI CRC 3-UTR CDK2-AP1 基因 T8 微卫星的新突变

• 批准号: 7118479
• 负责人: THOMAS K WEBER
• 金额: $ 8.3万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-28 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7118479
• 关键词: cell line; gene mutation; genes; human

DESCRIPTION (provided by applicant): . The objective of this application is to define the frequency and role of a recently discovered del T alternation in poly(T)8 in the 3'-UTR of the Cyclin Dependent Kinase-2- Associated Protein-1 (CDK2-AP1) gene in Microsatellite unstable (MSI) colorectal cancer (CRC). Current literature recognizes two pathways lead to invasive CRC. Microsatellite stable (MSS) pathway is the most common pathway, seen in approximately 80% of CRC cases, in which no microsatellite sequence alterations were observed. The other pathway is MSI pathway, explained in the remaining 20% of CRC cases, characterized by frame shift mutations and base-pair substitutions of the microsatellite DNA. CDK2-AP1 is a known growth suppressor in the etiology of RC. We previously reported that the CDK2-AP1 expression has been significantly decreased in human MS RC cell lines (Yuan et al., 2003) and its induction resulted in decreased cell proliferation and increased apoptosis (Kent et al., 2004). Recently, we detected a frequent, novel single alteration, del T in poly (T)8 of the 3'-UTR of the CDK2-AP1 gene in 25% of MSI CRC cell lines (3 of 12), is associated with decreased DK2-AP1 expression (Yuan et al., 2005). In a concurrent study by Ruggiero, described a similar poly T(8) alteration in 3'-UTR region of the CEACAM1 gene in human MSI CRC, strengthens our project. Based on our previously published findings, we hypothesize: 1). del T in a poly (T)8 of the 3'-UTR of the CDK2-AP1 gene is a frequent alteration in MSI CRC, 2) observed del T is a somatic alteration, associated with decreased expression of CDK2-AP1. In this proposed project, we will test our hypothesis by completion of he following Specific Aims: Specific Aim 1: a) We will determine the frequency of the del T in a poly (T)8 of he 3'-UTR of the CDK2-AP1 gene in human MSI CRC; b) To further determine this alteration is a somatic or a germline mutation, we will compare the identified positive CRC sample with normal tissue pairs. Specific Aim 2: We will characterize the role of this del T alteration in decreased expression of CDK2-AP1 in human MSI CRC. Completion of our proposed Specific Aims will advance our knowledge about the mechanism of decreased CDK2-AP1 expression in MMR deficient CRC. At the completion of this proposed pilot study, will urther determine del T alteration in a large number of CRC patients to identify high risk population.

描述（由申请人提供）：本申请的目的是确定最近发现的在微卫星不稳定（MSI）结直肠癌（CRC）中细胞周期蛋白依赖性激酶-2-相关蛋白-1（CDK 2-AP 1）基因的3 '-UTR中的聚（T）8中的del T改变的频率和作用。目前的文献认为有两种途径导致侵袭性CRC。微卫星稳定（MSS）途径是最常见的途径，见于约80%的CRC病例，其中未观察到微卫星序列改变。另一种途径是MSI途径，在其余20%的CRC病例中解释，其特征在于微卫星DNA的移码突变和碱基对取代。CDK 2-AP 1是RC病因学中已知的生长抑制因子。我们先前报道了CDK 2-AP 1表达在人MS RC细胞系中显著降低（Yuan等人，2003）并且其诱导导致细胞增殖减少和凋亡增加（肯特等人，2004年）。最近，我们在25%的MSI CRC细胞系（12个中的3个）中检测到一种频繁的、新的单一改变，即CDK 2-AP 1基因的3 '-UTR的聚（T）8中的del T，其与DK2-AP 1表达降低相关（Yuan et al.，2005年）。在Ruggiero的同时研究中，描述了人MSI CRC中CEACAM 1基因的3 '-UTR区域中类似的多聚T（8）改变，加强了我们的项目。根据我们先前发表的研究结果，我们假设：1）。CDK 2-AP 1基因的3 '-UTR的聚（T）8中的del T是MSI CRC中的常见改变，2）观察到的del T是体细胞改变，与CDK 2-AP 1的表达降低相关。在这个提议的项目中，我们将通过完成以下具体目标来测试我们的假设：具体目标1：a）我们将确定人MSI CRC中CDK 2-AP 1基因的3 '-UTR的聚（T）8中的del T的频率; B）为了进一步确定这种改变是体细胞突变还是种系突变，我们将比较鉴定的阳性CRC样品与正常组织对。具体目标2：我们将描述这种缺失T改变在人MSI CRC中CDK 2-AP 1表达降低中的作用。完成我们提出的特定目标将推进我们对MMR缺陷CRC中CDK 2-AP 1表达降低机制的认识。在完成这项拟议的初步研究后，将进一步确定大量CRC患者中的del T改变，以确定高危人群。