CYSTATIN C EXPRESSION AND B16 MELANOMA METASTASIS

这个项目的长期目标是了解半胱氨酸的作用。 蛋白水解酶及其抑制物与肿瘤细胞转移。转移是 癌症死亡的主要原因和转移的一个主要特征 细胞是指它们通过蛋白分解事件侵入正常组织的能力。 这项提案将半胱氨酸蛋白酶抑制剂Cystatin C视为 一种潜在的抗转移药物。需要检验的假设是 胱抑素C抑制B16黑色素瘤细胞侵袭 半胱氨酸蛋白酶活性。这项建议的具体目标是： 1.构建小鼠半胱氨酸氨基转移酶C表达载体 金属硫蛋白启动子允许Cystatin C在细胞中过表达 对锌的反应。 2.检测两种组织中半胱氨酸氨基转移酶C的表达 未转染及表达载体转染的B16黑色素瘤细胞系 作为对锌的反应。 3.转B16黑色素瘤细胞的体外侵袭参数 用锌诱导和不用锌诱导转染型半胱氨酸氨基转移酶的测定 C。 胱抑素C的过表达将决定肿瘤细胞的哪一步 转移需要半胱氨酸蛋白酶活性才能发生。

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.