Melanoma Proteoglycan and Matrix Metalloproteinases

性状：（由申请人提供）基质表达增加 金属蛋白酶（MMPs）与许多肿瘤的进展有关 包括恶性黑素瘤。MMP级联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-MMP），也需要 侵袭性肿瘤细胞表达基质金属蛋白酶和粘附受体。进展 与MT 1-MMP表达增加有关， 其它可溶性MMP，包括MMP-2、MMP-1和MMP-13。MT 1-MMP可以激活 proMMP-2明胶酶，导致ECM组分的更快降解， proMMP-1胶原酶的活化。在目前的提案中，我们提出 许多侵袭性原发性黑色素瘤细胞表达MT3-MMP（a 与MT 1-MMP相关的跨膜MMP）。表面表达或MT3-MMP刺激 原代黑色素瘤细胞体外侵袭天然I型胶原凝胶的实验研究 并导致体外明胶分解活性增加和肿瘤生长加速 在皮下注射到免疫受损的小鼠中后的生长。MT3-MMP 介导的侵袭需要MMP-2和MMP-1，这表明MT3-MMP 在肿瘤细胞表面启动MMP激活级联反应。MT3-MMP 介导的人黑色素瘤侵袭需要表达黑色素瘤细胞 表面蛋白聚糖（MCSP），一种大的跨膜粘附受体， 与绝大多数人类黑色素瘤有关。MCSP核心蛋白的抑制 表达，或抑制硫酸软骨素（CS）的加入， 合成MCSP核心蛋白，抑制黑色素瘤侵袭和明胶溶解 这些细胞中的活性。黑色素瘤中MT3-MMP与MCSP共沉淀 提取物，这种共沉淀取决于CS的存在， MCSP核心蛋白。此外，重组MT3-MMP、MMP-2和MMP-1均结合 到CS偶联的珠粒。这些结果表明，MCSP可能有助于定位 和/或激活侵袭性黑素瘤细胞表面上的MMP级联。的 目前的建议有两个主要目标。首先，我们将进一步定义 MT3-MMP表面表达与proMMP-2活化的关系 或proMMP-1在黑素瘤侵袭中的作用。其次，我们提出检验假设 MCSP用于结合和/或改变这三种细胞的活化， 入侵促进蛋白酶。了解硫酸软骨素相互作用 与基质金属蛋白酶可能导致新的疗法，以抑制黑色素瘤的侵袭， 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.