Melanoma Proteoglycan and Matrix Metalloproteinases

描述：(申请人提供)增加矩阵的表达 金属蛋白酶(MMPs)与许多肿瘤的进展有关 包括恶性黑色素瘤。基质金属蛋白酶的激活受以下因素控制 细胞表面的事件，从膜型的表面表达开始 基质金属蛋白酶(MT-MMPs)及其相互作用 侵袭性肿瘤细胞表达MMPs和黏附受体。进步论 原发性黑色素瘤的发生与MT1-MMPs的表达增加有关 其他可溶性MMPs，包括MMP2、MMP1和MMP13。MT1-基质金属蛋白酶可激活 前基质金属蛋白酶-2明胶酶，导致细胞外基质成分更快的降解，并 激活原基质金属蛋白酶-1胶原酶。在目前的提案中，我们提出 大量侵袭性原发黑色素瘤细胞表达MT3-MMPa 与MT1-MMPs相关的跨膜MMPs)。表面表达或MT3-MMPs刺激 天然I型胶原凝胶体外侵袭原代黑色素瘤细胞的实验研究 并导致体外明胶溶解活性增加和加速肿瘤 皮下注射免疫缺陷小鼠后的生长。MT3-基质金属蛋白酶 介导性侵袭需要基质金属蛋白酶-2和基质金属蛋白酶-1，提示MT3-基质金属蛋白酶 启动肿瘤细胞表面的基质金属蛋白酶激活级联反应。最后，MT3-MMPs 人黑色素瘤侵袭需要黑色素瘤细胞的表达 表面蛋白多糖(MCSP)--一种大的跨膜黏附受体 与绝大多数人类黑色素瘤有关。MCSP核心蛋白的抑制作用 表达，或抑制添加硫酸软骨素(CS)到新生的 合成MCSP核心蛋白，抑制黑色素瘤侵袭和明胶溶解 这些细胞中的活动。黑色素瘤中MT3-MMPs与MCSP共沉淀的研究 萃取物，这种共沉淀依赖于CS在 MCSP核心蛋白。此外，重组MT3-MMP3、MMP2和MMP1均可结合 到壳聚糖偶联珠。这些结果表明，MCSP可能有助于本地化 和/或激活侵袭性黑色素瘤细胞表面的基质金属蛋白酶。这个 目前的提案有两个主要目标。首先，我们将进一步定义 MT3-基质金属蛋白酶表面表达与原基质金属蛋白酶-2活化的关系 原基质金属蛋白酶-1在黑色素瘤侵袭中的作用其次，我们建议对假设进行检验 该MCSP用于绑定和/或修改这三个参数的激活 促进侵袭的蛋白酶。了解硫酸软骨素相互作用 MMPs可能会导致抑制黑色素瘤侵袭和 转移。

DESCRIPTION: (provided by applicant) Increased expression of matrix metalloproteinases (MMPs) is associated with the progression of many tumors including malignant melanoma. The activation of MMP cascades is controlled by events at the cell surface, starting with surface expression of membrane type matrix metalloproteinases (MT-MMP) and also requiring the interactions between MMPs and adhesion receptors expressed in invasive tumor cells. The progression of primary melanomas is associated with increased expression of MT1-MMP and other soluble MMPs, including MMP-2, MMP-1 and MMP-13. MT1-MMP can activate proMMP-2 gelatinase, leading to more rapid degradation of ECM components and to activation of proMMP-1 collagenase. In the current proposal, we present evidence that numerous invasive primary melanoma cells express MT3-MMP (a transmembrane MMP related to MT1-MMP). Surface expression or MT3-MMP stimulates invasion of primary melanoma cells through native type I collagen gels in vitro and leads to increased in vitro gelatinolytic activity and accelerated tumor growth following subcutaneous injections into immunocompromised mice. MT3-MMP mediated invasion requires both MMP-2 and MMP-1, suggesting that MT3-MMP initiates an MMP activation cascade at tumor cell surfaces. Finally, MT3-MMP mediated human melanoma invasion requires the expression of Melanoma Cell Surface Proteoglycan (MCSP), a large transmembrane adhesion receptor associated with the vast majority of human melanomas. Inhibition of the MCSP core protein expression, or inhibiting the addition of chondroitin sulfate (CS) to newly synthesized MCSP core protein, inhibits melanoma invasion and gelatinolytic activity in these cells. MT3-MMP co-precipitates with MCSP in melanoma extracts, and this co-precipitation is dependent on the presence of CS on the MCSP core protein. Furthermore, recombinant MT3-MMP, MMP-2 and MMP-1 all bind to CS-coupled beads. These results suggest that MCSP may help to localize and/or activate MMP cascades on the surface of invasive melanoma cells. The current proposal has two major goals. First we will further define the relationship between surface expression of MT3-MMP and activation of proMMP-2 or proMMP-1 in melanoma invasion. Secondly, we propose to test the hypothesis that MCSP serves to bind and/or modify the activation of these three invasion-promoting proteases. Understanding chondroitin sulfate interactions with MMPs may lead to new therapies to inhibit melanoma invasion and metastasis.