The role of SEPT9 in cell proliferation and oncogenesis

SEPT9在细胞增殖和肿瘤发生中的作用

基本信息

批准号：
7330309
负责人：
ELIZABETH M. PETTY
金额：
$ 29.6万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-09-08 至 2010-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7330309
关键词：
Abnormal Cell Agar Allelic Imbalance Anchorage-Independent Growth Aneuploidy Animals Apoptosis Apoptotic Breast Breast Adenocarcinoma Breast Cancer Cell Cell Cycle Cell Cycle Progression Cell Cycle Regulation Cell Nucleus Cell Proliferation Cell division Cells Cellular Morphology Characteristics Chromosome Band Chromosome Banding Clinical Clinical Management Colon Carcinoma Condition Contact Inhibition Coupled Cytokinesis Data Depth Development Ectopic Expression Epithelial Cells Exons Family GTP Binding Genetic Genome Stability Genomic Instability Giant Cells Gland Goals Growth Homologous Gene Human Image Immunocompromised Host In Vitro Kinetics Lead Life Literature Localized Malignant - descriptor Malignant Neoplasms Malignant neoplasm of ovary Mammalian Cell Mammary Neoplasms Mammary Tumorigenesis Mammary gland Microfilaments Modeling Molecular Morphology Mouse Mammary Tumor Virus Mus Mutagenesis Names Nuclear Nutrient Oncogenes Oncogenic Orthologous Gene Outcome Pathway interactions Patients Pattern Phenotype Play Ploidies Protein Family Protein Isoforms Protein Overexpression Proteins Proto-Oncogenes RNA Splicing Rate Regulation Research Personnel Role S-Phase Fraction SCID Mice Sampling Studies Small Interfering RNA Somatic Cell T-Cell Lymphoma T-Lymphocyte Testing Tetanus Helper Peptide Therapeutic Time Tissues Transcript Transgenes Transgenic Model Transgenic Organisms Tumorigenicity Variant Yeasts alpha Tubulin cancer cell carcinogenesis clinically significant immunocytochemistry in vitro Model in vivo innovation insight intracellular protein transport leukemia malignant breast neoplasm malignant phenotype member mouse model novel ovarian neoplasm pro-apoptotic protein prognostic programs promoter protein localization location tumor tumorigenesis

项目摘要

A classic feature of carcinogenesis is aberrant cell cycle regulation but little is known about how abnormal cell division contributes to malignant progression. Human SEPT9 belongs toa highly conserved eukaryotic family of GTP binding septin proteins importantin cell division. In yeast, septins are coupled to cell cycle progression and are critical for normal morphology. Mammalian SEPT9 is requiredfor normal somatic cell division. SEPT9 is highly homologous to Sept9/Sint1,a murineproto-oncogeneinvolvedin T-cell lymphomasthat is overexpressed in murine models of mammary adenocarcinomas. We originally cloned SEPT9 from a region of allelic imbalance in breast cancers and have subsequently demonstrated SEPT9 amplification in breast cancer cells. We have also observed preferential expression of an alternatively spliced variant, SEPT9v1, in breast cancer cells. Stable ectopic expression of SEPT9v1 studied to date in in vitro models resulted in increased growth kinetics, altered cell morphology, increasedfoci formation, increased invasiveness,abnormal cell division, nuclear mis-localization of the protein, and aneuploidy. SEPT9 was also identified as an MIL fusion partner in leukemia cells. Taken together, findings of: (1) an essential role for SEPT9 in cell division; (2) SEPT9 amplification and preferential expression of SEPT9v1 in breast cancer cells; (3) an altered cell phenotype with increased proliferation, invasiveness, foci formation, and aneuploidy when SEPT9v1variant is ectopically expressed; and (4) homology to an oncogenic murine ortholog overexpressedin breast cancer provide compelling support for our hypothesis that increased expression of SEPT9v1, and possibly additional SEPT9 variants, is associated with abnormal cell division and malignant progression in mammary epithelial cells. To test this hypothesis we will: (1) determine if cellular levels of SEPT9v 1 in an archival panel of well-characterized primary and metastatic mammary tumors correlate with poor prognostic clinicopathological variables clinical outcome; (2) analyze in vitro epithelial cell culture and in vivo tumorigenicity models to determine how increased SEPT9v1impacts cell proliferation, growth characteristics, invasiveness, apoptosis, cell morphology, and genomic stability as related to malignant progression; and (3) evaluate how in vivo overexpression of SEPT9v1 impacts normal mammary gland development and malignant potential by constructing and analyzing tissue specific and conditional transgenic mouse models, including crosses to other transgenicmurine modelsof breastcancer. Resultswill provide insights into how increased SEPT9 v1 expression impacts cell division and tumorigenesis in mammary epithelial cells, which may have significant prognostic and/or therapeutic implications.

癌变的一个经典特征是异常细胞周期调节，但对异常的了解鲜为人知细胞分裂有助于恶性进展。人类sept9属于TOA高度保守的真核家族 GTP结合的SEPTIN蛋白质的重要细胞分裂。在酵母中，septins耦合到细胞周期的进程，对于正常形态至关重要。哺乳动物SEPT9是正常的体细胞分裂所必需的。 sept9高度与sept9/sint1同源，鼠 - 互联素Volvedin t-cell淋巴瘤在鼠中过表达乳腺腺癌的模型。我们最初从乳房中等位基因失衡的地区克隆了sept9 癌症，随后证明了乳腺癌细胞中的SEPT9扩增。我们也有观察到在乳腺癌细胞中观察到的剪接变体Sept9v1的优先表达。稳定的迄今为止在体外模型中研究的SEPT9V1的异位表达导致生长动力学增加，改变了细胞形态，增加的形成，增加侵入性，异常细胞分裂，核误局化蛋白质和非整倍。 SEPT9也被确定为白血病细胞中的MIL融合伴侣。拍摄共同的发现：（1）SEPT9在细胞分裂中的重要作用；（2）SEPT9扩增和优先 SEPT9V1在乳腺癌细胞中的表达；（3）改变的细胞表型，增殖增加，当sept9v1variant在异位表达时，侵入性，焦点形成和非整倍性；（4）同源性致癌鼠直源性过表达乳腺癌为我们的假设提供了令人信服的支持 sept9v1的表达增加，可能还有其他SEPT9变体，与异常细胞有关乳腺上皮细胞的分裂和恶性进展。要检验该假设，我们将：（1）确定是否是否在特征良好的原发性和转移性乳腺肿瘤的档案板中的sept9v 1的细胞水平与预后不良的临床病理变量临床结果相关；（2）分析体外上皮细胞培养和体内肿瘤性模型，以确定升高的SEPT9V1如何影响细胞增殖，生长与恶性有关的特征，侵入性，凋亡，细胞形态和基因组稳定性进展；（3）评估SEPT9V1体内过度表达如何影响正常乳腺通过构建和分析组织特异性和条件转基因的发展和恶性潜力小鼠模型，包括乳腺癌其他转基因模型的交叉。 Resultsswill提供见解 sept9 v1表达增加如何影响乳腺上皮细胞的细胞分裂和肿瘤发生，，可能具有重大的预后和/或治疗意义。