The Role of Myopodin in Invasive Prostate Cancers

Myopodin 在侵袭性前列腺癌中的作用

基本信息

批准号：
7229025
负责人：
JIANHUA LUO
金额：
$ 24.98万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-07-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7229025
关键词：
4q25 Actins Allelotyping Anchorage-Independent Growth Antisense Oligonucleotides Apoptosis Appendix Behavior Binding Biological Biological Assay Biological Process Bundling Cell Line Cells Chromosomes Clinical Co-Immunoprecipitations Cytogenetics Cytoskeleton DNA Sequence DU145 Databases Deletion Mutation Detection Diagnostic Neoplasm Staging Disease Elderly Epithelium Etiology Exhibits Expressed Sequence Tags Fluorescent in Situ Hybridization Frequencies Gene Expression Genes Genetic Genome Genomics Gleason Grade for Prostate Cancer Heterogeneity Hormonal Hot Spot Human Immigration In Situ Hybridization In Vitro Indolent Inherited Invasive Kidney LNCaP Laboratories Laboratory Study Learning Life Light Loss of Heterozygosity Malignant Neoplasms Malignant neoplasm of prostate Maps Mediating Messenger RNA Metalloproteases Methodology Mitosis Molecular Abnormality Morphology Mus Muscle Cells Names Neoplasm Metastasis Neurons Normal Cell Northern Blotting Numbers Nutritional Oligonucleotides PC3 cell line PSA screening Pathogenesis Patients Pattern Penetrance Physiological Play Polymerase Chain Reaction Population Positioning Attribute Prostate Prostate Adenocarcinoma Prostate carcinoma Protein Overexpression Proteins Rate Regulation Relapse Relative (related person)Research Personnel Respiratory Diaphragm Reverse Transcriptase Polymerase Chain Reaction Role Sequence Analysis Series Site Southern Blotting Surveys System Testing Trisomy Western Blotting Wound Healing Y Chromosome Yeasts ZYX gene actin 2 autosomal dominant trait base cancer cell cell motility cell transformation chromosome X loss clinically relevant comparative genomic hybridization genetic regulatory protein in vivo indexing matrigel mortality mutant myopodin novel outcome forecast podocyte polymerization programs research study serum PSA size synaptopodin tumor tumor growth yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Prostate cancer is a disease with considerable biological heterogeneity and clinical aggressiveness. Although prostate cancer is prevalent among elderly population, only a small proportion of prostate cancer become invasive, metastatic and life-threatening. Thus, identifying genes and the underlying mechanisms that convert prostate cancer from a relatively indolent disease to an aggressive one hold the key for reducing the mortality of the disease. Recently, by using "differential subtraction chain", we identified a novel gene named "myopodin" that is frequently deleted in aggressive type of prostate cancer. Regardless of Gleason grade, deletions of myopodin, either partial or complete, correlate with high rate of invasion, metastasis or clinical relapse of prostate cancer. Sequence analysis indicates that myopodin shares significant homology with synaptopodin, a protein expressed in neurons and kidney podocytes and responsible for forming physiological cell-cell contact for these cells. Interestingly, over 90% of the partial deletions of myopodin are located in the region sharing homology with synaptopodin. Overexpression of myopodin in PC-3 or LNCaP cells reduced the invasiveness of these cell lines by 3 fold or more in matrigel traverse analysis, and by 10 fold in tumor growth in vivo. Overexpression of myopodin induces actin bundling and retards migration in PC-3. Yeast two-hybrid and co-immunoprecipitation analyses indicate that myopodin binds zyxin, a critical regulatory protein in actin cytoskeleton reorganization, cell motility and mitosis. In our proposal, we hypothesize that myopodin plays an important role in regulating cell motility, maintaining the integrity of normal cell-cell contacts of prostate epithelium, and inhibits the invasiveness of prostate cancer cells, and propose the following specific aims: 1) to overexpress myopodin in PC-3, Du145 and LNCaP cells and subsequently to test the changes of invasiveness of these cells through anchorage independent growth assay, matrigel traverse analysis, diaphragm invasion analysis, metastasis assay and metalloproteinase assays; 2) to define the motifs and minimal domain of myopodin essential for regulation of invasion by constructing arrays of deletion mutants and test their ability to regulate invasiveness; 3) to identify the functional significance of myopodin/zyxin interaction by focusing on cell motility and cytoskeleton re-organization related analysis on myopodin transformed cells; and 4) To identify potential effector genes that might mediate the biological function of myopodin through a comprehensive analysis of gene expression of myopodin expressing cells.

描述（申请人提供）：前列腺癌是一种具有相当大的生物学异质性和临床侵袭性的疾病。尽管前列腺癌在老年人群中普遍存在，但只有一小部分前列腺癌具有侵袭性、转移性和危及生命的能力。因此，识别将前列腺癌从一种相对惰性的疾病转变为侵袭性疾病的基因和潜在机制是降低该疾病死亡率的关键。最近，通过使用“差异消减链”，我们鉴定了一种名为“myopodin”的新基因，该基因在侵袭性前列腺癌中经常被删除。无论格里森分级如何，肌足蛋白的部分或完全缺失都与前列腺癌的高侵袭、转移或临床复发率相关。序列分析表明，肌足蛋白与突触足蛋白具有显着的同源性，突触足蛋白是一种在神经元和肾足细胞中表达的蛋白质，负责形成这些细胞的生理细胞间接触。有趣的是，超过 90% 的肌足蛋白部分缺失位于与突触足蛋白具有同源性的区域。在基质胶遍历分析中，PC-3 或 LNCaP 细胞中肌足蛋白的过度表达使这些细胞系的侵袭性降低了 3 倍或更多，在体内肿瘤生长中使这些细胞系的侵袭性降低了 10 倍。 Myopodin 的过度表达会诱导肌动蛋白成束并延迟 PC-3 中的迁移。酵母双杂交和免疫共沉淀分析表明，myopodin 与 zyxin 结合，zyxin 是肌动蛋白细胞骨架重组、细胞运动和有丝分裂中的关键调节蛋白。在我们的提议中，我们假设Myopodin在调节细胞运动、维持前列腺上皮正常细胞-细胞接触的完整性以及抑制前列腺癌细胞的侵袭方面发挥重要作用，并提出以下具体目标：1）过表达PC-3、Du145和LNCaP细胞中的myopodin，随后通过贴壁独立生长测定、基质胶横移分析、膈肌侵袭分析来测试这些细胞的侵袭性变化，转移测定和金属蛋白酶测定； 2）通过构建缺失突变体阵列来定义调控入侵所必需的myopodin的基序和最小结构域并测试其调控入侵的能力； 3）通过对Myopodin转化细胞的细胞运动和细胞骨架重组相关分析来确定Myopodin/zyxin相互作用的功能意义； 4)通过对Myopodin表达细胞的基因表达进行综合分析，鉴定可能介导Myopodin生物学功能的潜在效应基因。