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。

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.