Role of Cdc2 in HPV E6-mediated Apoptosis

Cdc2 在 HPV E6 介导的细胞凋亡中的作用

基本信息

批准号：
7081360
负责人：
JASON J CHEN
金额：
$ 7.93万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-01 至 2007-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7081360
关键词：
RNA interference apoptosis cell cycle proteins cyclin dependent kinase enzyme activity gene mutation human papillomavirus keratinocyte tissue /cell culture virus genetics virus replication

项目摘要

DESCRIPTION (provided by applicant): Infection with the human papillomaviruses (HPV) induces warts and is strongly associated with the development of cervical cancer. Our long-term goal is to elucidate the molecular basis for HPV-induced lesions and to develop therapeutic strategies for the elimination of HPV infection, Apoptosis is a genetically programmed process of cellular destruction that can eliminate virally infected cells or tumors. Viruses such as HPV have evolved specific gene products to modulate apoptosis. Induction of apoptosis is a desirable approach in treating HPV-induced lesions. The HPV E6 gene is essential for viral replication and induces apoptosis. However, the mechanism by which E3 mediates cellular sensitization to apoptosis remains to be determined. To understand the molecular basis for E6-mediated apoptosis, we have established human keratinocytes expressing HPV E6 and found these cells to be susceptible to apoptosis induced by chemotherapeutic agents. To identify genes responsible for E6-mediated apoptosis, a proteomic approach was taken. Significantly, the level and activity of Cdc2, the cyclin-dependent kinase required for cells to enter mitosis and whose aberrant activation has been associated with apoptosis, was increased in E6-expressing cells undergoing apoptosis. An E6 mutant that is unable to activate Cdc2 is defective in sensitizing cells to apoptosis. Moreover, inhibition of Cdc2 activity with specific inhibitor reduced ap6ptosis. We hypothesize that activation of Cdc2 plays an important role in E6-mediated cellular sensitization to apoptosis. The specific aim of this proposal is designed to test this possibility with the following approaches: (1) to examine the relevance of Cdc2 to E6-mediated cellular sensitization to apoptosis through a genetic approach and (2) to determine the role of Cdc2 in E6-mediated apoptosis by molecular ablation and ectopic expression. These studies will help understand the mechanism of E6-mediated apoptosis and contribute to the development of therapeutic approaches against HPV-induced diseases.

描述（由申请人提供）：对人乳头瘤病毒（HPV）的感染会诱导疣，并且与宫颈癌的发展密切相关。我们的长期目标是阐明HPV诱导的病变的分子基础，并制定消除HPV感染的治疗策略，凋亡是一种基因编程的细胞破坏过程，可以消除病毒感染的细胞或肿瘤。诸如HPV之类的病毒已进化出特定的基因产物以调节细胞凋亡。凋亡诱导是治疗HPV诱导的病变的理想方法。 HPV E6基因对于病毒复制至关重要，并诱导凋亡。但是，E3介导细胞敏化对凋亡的机制仍有待确定。为了了解E6介导的凋亡的分子基础，我们已经建立了表达HPV E6的人角质形成细胞，并发现这些细胞易受化学治疗剂诱导的凋亡。为了鉴定负责E6介导的细胞凋亡的基因，采取了一种蛋白质组学方法。值得注意的是，在经历凋亡的E6表达细胞中增加了Cdc2的水平和活性，即细胞进入有丝分裂所需的细胞周期蛋白依赖性激酶，其异常激活与凋亡相关。无法激活CDC2的E6突变体在使细胞对凋亡敏化时有缺陷。此外，用特异性抑制剂降低AP6PTOSIS的抑制Cdc2活性。我们假设Cdc2的激活在E6介导的细胞对凋亡的敏感性中起重要作用。该提案的具体目的旨在通过以下方法测试这种可能性：（1）通过遗传方法检查Cdc2与E6介导的细胞敏化与凋亡的相关性，以及（2）确定Cdc2在E6介导的凋亡中通过分子烧蚀和分解表达的作用。这些研究将有助于了解E6介导的凋亡的机制，并有助于开发针对HPV诱导疾病的治疗方法。