MODULATION OF TYPE-IV COLLAGENASE ACTIVITY AND INVASIVE BEHAVIOR OF METASTATIC HUMAN-MELANOMA (A2058) CELLS-INVITRO BY MONOCLONAL-ANTIBODIES TO TYPE-IV COLLAGENASE
MODULATION OF TYPE-IV COLLAGENASE ACTIVITY AND INVASIVE BEHAVIOR OF METASTATIC HUMAN-MELANOMA (A2058) CELLS-INVITRO BY MONOCLONAL-ANTIBODIES TO TYPE-IV COLLAGENASE
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DOI：
10.1002/ijc.2910460224
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发表时间：
1990-08-15
影响因子：
6.4
通讯作者：
TRYGGVASON, K
中科院分区：
文献类型：
--
作者：
HOYHTYA, M;HUJANEN, E;TRYGGVASON, K
制备了抗人iv型胶原酶的单克隆抗体（mab），并用于酶活性和肿瘤细胞侵袭的体外研究。从人黑色素瘤细胞（A2058）无血清培养基中纯化的酶产生了15个单抗克隆。选取影响ⅳ型胶原酶活性的5个克隆进行进一步鉴定。所有选择的克隆均可用IgG-Sepharose亲和柱一步纯化iv型胶原酶。当3h -脯氨酸标记的iv型胶原作为底物时，其中一种抗体激活了酶。活化依赖于酶抗体比。4个无性系的活性抑制率均在30%以上，最大抑制率为50%。有趣的是，在体外入侵实验中，激活该酶的同一抗体也通过重组基膜增加了对A2058细胞的入侵。4种抑制抗体降低了A2058细胞通过重组基膜的渗透。这些结果有力地支持了先前关于iv型胶原酶在肿瘤细胞侵袭中的重要性的发现。
Monoclonal antibodies (MAbs) against human type-IV collagenase were developed and used for studies on enzyme activity and tumor-cell invasion in vitro. Fifteen MAb clones were generated against the enzyme purified from serum-free culture medium of human melanoma cells (A2058). Five clones affecting the activity of type-IV collagenase were selected for further characterization. All the selected clones could be used for a single-step purification of type-IV collagenase using IgG-Sepharose affinity columns. One of the antibodies activated the enzyme when 3H-proline-labelled type-IV collagen was used as substrate. The activation was dependent on the enzyme antibody ratio. Four clones more than 30% inhibition of the activity, maximal inhibition being 50%. Interestingly, the same antibody which activated the enzyme also increase the invasion of A2058 cells through a reconstituted basement membrane in an in vitro invasion assay. The 4 inhibitory antibodies decreased the penetration of A2058 cells through the reconstituted basement membrane. The results strongly support previous findings about the importance of type-IV collagenase in tumor-cell invasion.