权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

CYSTATIN C EXPRESSION AND B16 MELANOMA METASTASIS

胱抑素 C 表达与 B16 黑色素瘤转移

基本信息

批准号：
2105392
负责人：
JAMES Lewis COX
金额：
$ 11.54万
依托单位：
A.T. STILL UNIVERSITY OF HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-08-01 至 1998-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2105392
关键词：
animal genetic material tag complementary DNA electrophoresis enzyme activity enzyme inhibitors gene expression genetic promoter element growth inhibitors melanoma metallothionein metastasis plasmids protein purification tissue /cell culture transfection zinc

项目摘要

The long-term goal of this project is to understand the role of cysteine proteinases and their inhibitors in tumor cell metastasis. Metastasis is the major cause of cancer mortality and a dominant feature of metastatic cells is their ability to invade normal tissues via proteolytic events. This proposal will look at the cysteine proteinase inhibitor cystatin C as a potential anti-metastatic agent. The hypothesis to be tested is that cystatin C can block invasion by B16 melanoma cells due to inhibition of cysteine proteinase activity. The specific aims for this proposal are: 1. Creation of a mouse cystatin C expression plasmid driven by a metallothionein promoter that will permit overexpression of cystatin C in response to zinc. 2. Measurement of the expression levels of cystatin C in both untransfected and expression plasmid transfected B16 melanoma cell lines in response to zinc. 3. In vitro invasion parameters for transfected B16 melanoma cells will be measured both with and without zinc induction of transfected cystatin C. Overexpression of cystatin C will determine which steps of tumor cell metastasis require cysteine proteinase activity to occur.

这个项目的长期目标是了解半胱氨酸的作用蛋白酶及其抑制剂在肿瘤细胞转移中的作用。转移是癌症死亡率的主要原因和转移性肿瘤的主要特征细胞通过蛋白水解事件侵入正常组织的能力。该提案将半胱氨酸蛋白酶抑制剂胱抑素C视为一种潜在的抗转移剂有待检验的假设是，半胱氨酸蛋白酶抑制剂C可以阻断B16黑色素瘤细胞的侵袭，半胱氨酸蛋白酶活性这项建议的具体目标是： 1. 由半胱氨酸蛋白酶抑制剂驱动的小鼠胱抑素C表达质粒的产生金属硫蛋白启动子，将允许半胱氨酸蛋白酶抑制剂C的过表达，对锌的反应 2. 测量两种细胞中胱抑素C的表达水平未转染和表达质粒转染的B16黑色素瘤细胞系对锌的反应。 3. 转染的B16黑素瘤细胞的体外侵袭参数将在有和没有锌诱导转染的半胱氨酸蛋白酶抑制剂的情况下测量 C. Cystatin C的过表达将决定肿瘤细胞的生长步骤，转移需要半胱氨酸蛋白酶活性发生。