Novel tumor suppressor gene discovery in pancreatic cancer

胰腺癌中新的抑癌基因的发现

基本信息

批准号：
7256591
负责人：
JAMES R. ESHLEMAN
金额：
$ 13.12万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2009-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7256591
关键词：
Abbreviations Agar Alleles Biology Cancer Etiology Cell Line Cells Cessation of life Clone Cells Colon Carcinoma Colorectal Cancer DNA Sequence Detection Diploid Cells Discipline Dominant-Negative Mutation Epigenetic Process Etiology Event Expressed Sequence Tags Fluorouracil Genes Genetic Genetic Screening Genome Genomic Library Growth Human Individual Infection Insertional Mutagenesis Knock-out Libraries MADH4 gene Malignant Neoplasms Malignant neoplasm of pancreas Malignant neoplasm of urinary bladder Mammalian Cell Messenger RNA Molecular Mutate Mutation Nude Mice Pancreas Polymerase Chain Reaction Prevention Prostate Proteins Publishing RNA RNA Interference Research Reverse Transcriptase Polymerase Chain Reaction Role Small Interfering RNA System TP53 gene Technology Testing Time Tumor Suppressor Genes Work carcinogenesis gene discovery innovation malignant mouth neoplasm model development new technology novel small hairpin RNA tool tumor tumor growth tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): Pancreatic cancer is notoriously lethal. A full understanding of the oncogenes and tumor suppressor genes mutated in this cancer will aide both diagnostically and therapeutically. Genetic screens in diploid cells have been difficult until recently. Despite the existence of powerful selection systems, insertional mutagenesis of one allele can only inactivate the second copy by producing a dominant negative gene product, a relatively rare event. Most tumor suppressor genes however require bi-allelic inactivation. RNA interference (RNAi) is a novel technology that can destroy a given species of mRNA in cells. Libraries are now commercially available that target all known human genes, and are ideal tools for gene discovery if one has a powerful selection system, such as tumorigenesis. This proposal uses this technology to identify novel pancreatic cancer tumor suppressor genes. Hypothesis: Novel pancreas cancer tumor suppressor genes can be functionally identified using whole genome RNAi libraries. SA1: Construct cell lines. Specifically, transduce non-tumorigenic and weakly- tumorigenic pancreas cell lines with whole-genome RNAi libraries. SA2: Select tumorigenic cell clones. Using growth in soft agar and tumor growth in nude mice, and select cell clones that have become tumorigenic. SA3: Sequence RNAi's and test independent RNAi's. Identify the responsible RNAi by PCR and sequencing, and confirm suspect tumor suppressor genes using independent RNAi constructs.

描述（由申请人提供）：众所周知，胰腺癌是致命的。充分了解这种癌症中突变的癌基因和抑癌基因将有助于诊断和治疗。直到最近，二倍体细胞的遗传筛选一直很困难。尽管存在强大的选择系统，一个等位基因的插入诱变只能通过产生显性失活基因产物来使第二个拷贝失活，这是一种相对罕见的事件。然而，大多数肿瘤抑制基因需要双等位基因失活。 RNA 干扰 (RNAi) 是一种新技术，可以破坏细胞中特定种类的 mRNA。现在商业上可以买到针对所有已知人类基因的文库，如果拥有强大的选择系统（例如肿瘤发生），那么它们是基因发现的理想工具。该提案利用该技术来鉴定新型胰腺癌抑癌基因。假设：使用全基因组 RNAi 文库可以功能性地鉴定新型胰腺癌肿瘤抑制基因。 SA1：构建细胞系。具体来说，用全基因组 RNAi 文库转导非致瘤性和弱致瘤性胰腺细胞系。 SA2：选择致瘤细胞克隆。利用软琼脂中的生长和裸鼠中的肿瘤生长，并选择已致瘤的细胞克隆。 SA3：对 RNAi 进行测序并测试独立的 RNAi。通过 PCR 和测序鉴定负责的 RNAi，并使用独立的 RNAi 构建体确认可疑的肿瘤抑制基因。