Assessing Matrix Metalloproteinase Expression and Activity in Hepatocellular Carcinomas

The matrix metalloproteinases (MMPs) constitute a large family of zincand calcium-dependent endopeptidases that cleave extracellular matrix components (1 ). Hence, MMPs are classified according to their substrate specificities: interstitial collagenases, stromelysins, gelatinases, membrane-type matrix metalloproteinases (MT-MMPs), and elastase (Table 1 ). The regulation of MMP activity involves gene expression, proteolytic processing of the propeptides to active forms, and inhibition by specific tissue inhibitors of matrix metalloproteinase (TIMPs). MMPs are involved in situations that require extracellular matrix remodeling, including wound healing, development, inflammation, fibrosis angiogenesis, and tumor invasion (2 –5 ). Several complementary methods have provided an insightful description of the expression levels of MMPs and their pathological correlates. These include immunohistochemistry and Northern, Western, and dot blots. Additionally, the activity of MMPs is evaluated by gel substrate analysis. This approach has demonstrated that an increase in the expression of MMP2 (6 ,7 ), MT1-MMP (7 ), TIMP1, and TIMP2 (8 –10 ) is associated with liver fibrosis. Similarly, in hepatocellular carcinomas, a high expression of MMP2, MMP9, MT1-MMP, and matrilysin is related to tumor aggressiveness (11 –14 ). Consistently, by gel substrate analysis, MMP2 activity is increased in primary and secondary liver cancers (13 ,15 ,16 ). By in situ hybridization, the sources of