Role of Polo-Like Kinase-1 in Melanocytic Transformation

Polo 样激酶 1 在黑素细胞转化中的作用

基本信息

批准号：
8492041
负责人：
Nihal Ahmad
金额：
$ 30.31万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8492041
关键词：
Affect American Joint Committee on Cancer Aneuploidy Apoptosis Binding Biological Biological Assay Cell Cycle Cell Cycle Progression Cell Death Cell Line Cell Proliferation Cell division Cell physiology Cells Centrosome Chemicals Chromosomal Instability Clinical Consensus Cutaneous Melanoma Data Development Disease Drosophila genus Dysplastic Nevus Exclusion Family Genetic Growth Histone H3 Homologous Gene Human Immunoblotting Immunoprecipitation Implant In Vitro Knowledge Luciferases Lymph Node Involvement Malignant Neoplasms Mediating Melanocytic nevus Melanoma Cell Mitosis Mitotic Molecular Normal Cell Notch Signaling Pathway Nude Mice Operative Surgical Procedures Outcome Patients Pattern Phenotype Phosphorylation Phosphorylation Site Phosphotransferases Play Process Protein-Serine-Threonine Kinases Radioactive Regulation Reporting Reverse Transcriptase Polymerase Chain Reaction Role S-Phase Fraction Sampling Sentinel Lymph Node Signal Transduction Site Site-Directed Mutagenesis Skin Tissue Specimen Staging Techniques Testing Therapeutic Time Tissues Trypan Blue Tumor Volume Tumor Weights Tumorigenicity Ubiquitination Xenograft Model Xenograft procedure annexin A5 cancer cell cyclin B1 human PLK1 protein in vivo indexing inhibitor/antagonist melanocyte melanoma member neoplastic notch protein novel novel strategies outcome forecast overexpression preclinical study prognostic public health relevance small hairpin RNA small molecule tissue culture tumor tumor growth tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): Understanding of the mechanisms of melanocytic transformation is a continuing challenge. An in- depth knowledge of the genetic controls of cellular proliferation and cell division may provide critical information regarding the conversion of a normal melanocyte to its neoplastic phenotype. Polo-like kinases (Plks) are a family of highly conserved mitotic serine/threonine kinases that have been shown to play critical roles in cell division and cycle progression. Plk1 is the most widely studied member of the Plk family that contributes to multiple mitotic processes including the activation of the Cdk1/Cyclin B1 cascade, centrosome maturation, bipolar spindle formation and the regulation of Emi1 degradation at mitotic exit. Plk1 has been shown to be over-expressed in several cancers and a forced over-expression of Plk1 in normal cells has been found to cause a transformed phenotype and increased tumorigenicity. However, the functional role of Plk1 in melanocytic transformation is not well understood. In a recent study (J Invest Dermatol, In Press; Appendix-1), we have found that, i) Plk1 was over-expressed in both clinical tissue specimens and cultured human melanoma cells when compared to normal skin tissues and cultured normal melanocytes, respectively; and ii) genetic as well as chemical inhibition of Plk1 resulted in a significant decrease in the viability and growth of melanoma cells without affecting the normal human melanocytes. This study suggested that Plk1 may have an important role in melanoma survival and/or progression. However, additional studies are needed to determine the functional significance of Plk1 in melanocytic transformation and to define its molecular mechanism(s). In this study, we propose to challenge our novel hypothesis that Plk1 plays a critical role in melanocytic transformation and melanoma survival through modulation of Notch signaling. To test the hypothesis, the following specific aims are proposed: 1) To determine the expression pattern of Plk1, Notch and Numb during melanocytic transformation and melanoma development; 2) To determine the involvement of Notch signaling as a downstream target of Plk1 in normal melanocytes and melanoma cells; 3) To define the mechanism by which Plk1 regulates Notch1 signaling; and 4) To determine the functional and therapeutic significance of Plk1 in vivo in athymic nude mice xenografts. The outcome of proposed studies may define i) the role of Plk1 in melanocytic transformation and melanoma progression, ii) a novel connection between a key mitotic regulator (Plk1) with an important signaling pathway (Notch) involved in multiple cellular processes, including melanoma development.

描述（由申请人提供）：了解黑素细胞转化的机制是一个持续的挑战。对细胞增殖和细胞分裂的遗传控制的深入了解可能会提供有关正常黑素细胞转化为肿瘤表型的关键信息。 polo样激酶（PLK）是一个高度保守的有丝氨酸/苏氨酸激酶的家族，这些家族已被证明在细胞分裂和循环进程中起着关键作用。 PLK1是PLK家族中研究最广泛的成员，它有助于多种有丝分裂过程，包括激活CDK1/Cyclin B1级联反应，中心体成熟，双极纺锤体形成以及在有丝分裂出口时EMI1降解的调节。已显示PLK1在几种癌症中被表达过表达，并且已发现正常细胞中PLK1的强制表达会导致转化的表型和增加的肿瘤性。但是，PLK1在黑素细胞转化中的功能作用尚不清楚。在最近的一项研究（J Invest Dermatol，在印刷中；附录1），我们发现，与正常的皮肤组织和培养的正常黑素细胞相比，i）PLK1在临床组织样本和培养的人黑色素瘤细胞中都过表达。 ii）遗传和化学抑制PLK1导致黑色素瘤细胞的生存力和生长显着降低，而不会影响正常的人黑色素细胞。这项研究表明，PLK1可能在黑色素瘤存活和/或进展中起重要作用。但是，需要进行其他研究来确定PLK1在黑素细胞转化中的功能意义并定义其分子机制。在这项研究中，我们建议挑战我们的新假设，即PLK1通过调节Notch信号传导在黑素细胞转化和黑色素瘤存活中起关键作用。为了检验假设，提出了以下特定目的：1）确定在黑色素细胞转化和黑色素瘤发育过程中PLK1，缺口和麻木的表达模式； 2）确定Notch信号作为PLK1的下游靶标在正常的黑色素细胞和黑色素瘤细胞中； 3）定义PLK1调节Notch1信号传导的机制； 4）确定plk1在无胸腺裸鼠异种移植物中体内的功能和治疗意义。拟议研究的结果可能定义了i）PLK1在黑色素细胞转化和黑色素瘤进展中的作用，ii）与多种细胞过程有关的重要信号传导途径（Notch）之间的关键有丝分裂调节剂（PLK1）之间的新联系，包括黑色素瘤发育。