Identification of activated matrix metalloproteinase-2 (MMP-2) as the main gelatinolytic enzyme in malignant melanoma by in situ zymography
Identification of activated matrix metalloproteinase-2 (MMP-2) as the main gelatinolytic enzyme in malignant melanoma by in situ zymography
复制标题
DOI：
10.1002/path.1080
复制
发表时间：
2002-06-01
影响因子：
7.3
通讯作者：
Mauch, C
中科院分区：
文献类型：
--
作者：
Kurschat, P;Wickenhauser, C;Mauch, C
细胞外基质降解酶，如基质金属蛋白酶和丝氨酸蛋白酶，在肿瘤进展和转移过程中的参与是众所周知的。特别地，已经显示通过膜结合的MTI-MMP激活恶性细胞表面上的基质金属蛋白酶原（MMP）-2有助于各种肿瘤的侵袭能力。这项研究提供的证据表明，在恶性黑色素瘤组织中，有效的明胶分解活性增加主要位于黑色素瘤细胞与周围细胞外基质相互作用的部位。本文应用明胶原位酶谱技术对41例原发性黑色素瘤（浅表扩散型30例，结节型11例）和6例淋巴结转移瘤进行了研究。该技术定位组织切片内的有效蛋白水解活性区域。在28/41（68%）原发性黑色素瘤和6/6（100%）转移瘤中，在肿瘤的侵袭部分，特别是在肿瘤-基质相互作用的部位检测到相当大的蛋白水解，而在肿瘤细胞的固体巢的中心内没有或只有弱的蛋白水解活性。对从显微切割的黑色素瘤标本的不同区域获得的提取物进行酶谱分析，发现活化的MMP-2是负责这种活性的酶。免疫组化分析检测到MMP-2和MT 1-MMP的强染色，即使在没有发现蛋白水解活性的地区，通过原位酶谱，强调了更多的功能技术的重要性，平衡的蛋白水解系统的调查。该技术使得可以得出关于活化的蛋白酶和抑制剂之间的平衡的结论，这些蛋白酶和抑制剂经常被发现在体内紧密接近地一起存在。版权所有（C）2002约翰·维利父子有限公司
The involvement of extracellular matrix-degrading enzymes, such as matrix metalloproteinases and serine proteases, during tumour progression and metastasis is well established. In particular, the activation of pro-matrix metalloproteinase (MMP)-2 on the surface of malignant cells by membrane-bound MTI-MMP has been shown to contribute to the invasive abilities of various tumours. This study presents evidence that in tissue of malignant melanomas, increased effective gelatinolytic activity is mainly located at sites where melanoma cells interact with the surrounding extracellular matrix. Forty-one primary melanomas (30 superficial spreading and 11 nodular type) and six lymph node metastases were investigated by a modified technique of gelatin in situ zymography. This technique localizes areas of effective proteolytic activity within tissue sections. In 28/41 (68%) primary melanomas and in 6/6 (100%) metastases, considerable proteolysis was detected at the invading part of the tumour and especially at sites of tumour-stroma interactions, whereas no or only weak proteolytic activity was localized within the centres of solid nests of tumour cells. Zymographic analysis of extracts obtained from different areas of microdissected melanoma specimens identified activated MMP-2 as the enzyme responsible for this activity. Immunohistochemical analysis detected strong staining for MMP-2 and MT1-MMP, even in areas in which no proteolytic activity was found by in situ zymography, emphasizing the importance of more functional techniques for the investigation of balanced proteolytic systems. This technology makes it possible to draw conclusions regarding the balance between activated proteases and inhibitors, which are frequently found to be present together in close proximity in vivo. Copyright (C) 2002 John Viley Sons, Ltd.