Characterization of gelatinases linked to extracellular matrix invasion in ovarian adenocarcinoma: purification of matrix metalloproteinase 2.
Characterization of gelatinases linked to extracellular matrix invasion in ovarian adenocarcinoma: purification of matrix metalloproteinase 2.
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与卵巢腺癌细胞外基质侵袭相关的明胶酶的表征：基质金属蛋白酶 2 的纯化。
DOI：
10.1006/gyno.1996.0195
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发表时间：
1996
影响因子：
4.7
通讯作者：
Stack,MS
中科院分区：
文献类型：
--
作者：
Young,TN;Rodriguez,GC;Rinehart,AR;BastJr,RC;Pizzo,SV;Stack,MS
大量证据表明，细胞外基质的蛋白降解是癌细胞侵袭和转移所必需的。我们以前的工作已经证明了培养的卵巢腺癌细胞分泌两种明胶溶解金属蛋白酶，一种类似于基质金属蛋白酶2(MMP2)的72 kDa的酶和一种类似于MMP9的92 kDa的酶(Moseret al.，Int.J.Cancer56,552-559,1994)。为了评估这些酶的潜在活性相关性，我们使用明胶底物酶谱技术检测了卵巢癌腹水。在大多数腹水中发现了与几个卵巢癌细胞株分泌的基质金属蛋白酶相同的种类：基质金属蛋白酶-2样明胶酶(23/23例)和基质金属蛋白酶-9-样明胶酶(18/23例)，这表明这些种类在卵巢癌微环境中普遍存在，并可用于肿瘤相关的蛋白分解。通过检测肿瘤细胞介导的天然内皮细胞外基质(ECM)的蛋白分解和肿瘤细胞对重组基底膜(Matrigel)的侵袭，进一步证实了这些蛋白在卵巢癌侵袭中的作用。这些数据表明，一系列独立分离的卵巢腺癌细胞株分泌的IV型胶原酶活性与这些细胞分解ECM和侵袭基底膜的能力密切相关。此外，我们还鉴定并鉴定了卵巢癌相关的明胶酶，即在DOV 13细胞系的条件培养液中发现的72 kDa的明胶酶，称为基质金属蛋白酶-2。通过Western印迹、氨基末端序列和底物特异性分析，该酶与先前从其他来源获得的基质金属蛋白酶-2完全相同。此外，在DOV 13条件培养液中发现的很大一部分MMP-2活性在没有有机汞处理的情况下是活跃的，这表明卵巢癌细胞具有内源性酶原激活剂。综上所述，肿瘤相关蛋白水解酶介导的ECM蛋白降解在卵巢癌的侵袭和/或转移中起重要作用。
Substantial evidence indicates that proteolytic degradation of the extracellular matrix is necessary for invasion and metastasis by cancer cells. Our previous work has demonstrated elevated secretion by cultured ovarian adenocarcinoma cells of two gelatinolytic metalloproteinases, a 72-kDa enzyme resembling matrix metalloproteinase 2 (MMP-2) and a 92-kDa enzyme resembling MMP-9 (Moseret al., Int. J. Cancer56, 552–559, 1994). To assess the potentialin vivorelevance of these enzymes, we have examined ovarian carcinoma ascites using gelatin substrate zymography. MMP species identical to those secreted from several well-characterized ovarian adenocarcinoma cell lines were found in the majority of ascites: MMP-2-like gelatinase (23 of 23 cases) and MMP-9-like gelatinase (18 of 23 cases), suggesting a prevalence of these species in the ovarian carcinoma microenvironment and their availability for tumor-associated proteolysis. The contribution of these proteinases to ovarian cancer invasion was further demonstrated by experiments measuring tumor cell-mediated proteolysis of native endothelial cell extracellular matrix (ECM) and tumor cell invasion of reconstituted basement membrane (Matrigel). These data showed that secretion of type IV collagenase activity by a series of independently isolated ovarian adenocarcinoma cell lines correlated well with the ability of these cells to proteolyze the ECM and invade the basement membrane. Furthermore, we have identified and characterized an ovarian carcinoma-associated gelatinase, the 72-kDa MMP found in conditioned media of the DOV 13 cell line, as MMP-2. This enzyme was identical to the previously described MMP-2 from other sources by Western blot, amino terminal sequence, and substrate specificity. Additionally, a large portion of the MMP-2 activity found in DOV 13 conditioned media is active without organomercurial treatment, suggesting that ovarian cancer cells have an endogenous activator of the zymogen. Together, these data suggest that ECM proteolysis mediated by tumor-associated proteinases plays an important role in the invasion and/or metastasis of ovarian carcinoma.