Zhou H, Bernhard E J, Fox F E, Billings P C
Department of Radiation Oncology, University of Pennsylvania School of Medicine, Philadelphia 19104.
Biochim Biophys Acta. 1993 Jun 6;1177(2):174-8. doi: 10.1016/0167-4889(93)90037-p.
Matrix metalloproteinases are thought to play major roles in a wide array of normal and pathological processes. These proteinases are involved in the degradation of the extracellular matrix and are believed to facilitate the movement of cells from one site to another. In the current study, we examined the expression of the 92 kDa gelatinase activity (MMP-9) by the human T-lymphoma cell line, HSB. Proteinase activity was greatly elevated when cells were treated with TPA. This induction was initially observed at 6 h post-TPA treatment and continued to increase up to 48 h. Proteinase induction was inhibited by actinomycin D and cycloheximide, indicating that nascent RNA and protein synthesis were required. Staurosporine, an inhibitor of protein kinase C activity, suppressed the TPA-induction of gelatinase activity. Our results suggest that TPA induces the 92 kDa gelatinase activity by activating protein kinase C. TGF-beta also induced proteinase activity, although to a lesser extent than TPA. Several criteria indicate that this enzyme is a member of the family of matrix metalloproteinases: (1) this activity was inhibited by EDTA, 1,10-phenanthroline and TIMP; (2) this activity bound to a gelatin-agarose affinity resin; (3) it has a mass of approx. 92 kDa on SDS-polyacrylamide gels; (4) it cleaves gelatin and (5) the inducible proteinase cross reacts with antiserum to MMP-9.
基质金属蛋白酶被认为在一系列正常和病理过程中发挥主要作用。这些蛋白酶参与细胞外基质的降解，并被认为有助于细胞从一个部位移动到另一个部位。在本研究中，我们检测了人T淋巴瘤细胞系HSB中92 kDa明胶酶活性（MMP-9）的表达。用佛波酯（TPA）处理细胞时，蛋白酶活性显著升高。这种诱导最初在TPA处理后6小时观察到，并持续增加至48小时。放线菌素D和环己酰亚胺抑制蛋白酶诱导，表明需要新生RNA和蛋白质合成。蛋白激酶C活性抑制剂星形孢菌素抑制了TPA诱导的明胶酶活性。我们的结果表明，TPA通过激活蛋白激酶C诱导92 kDa明胶酶活性。转化生长因子-β（TGF-β）也诱导蛋白酶活性，尽管程度低于TPA。几个标准表明这种酶是基质金属蛋白酶家族的一员：（1）这种活性被乙二胺四乙酸（EDTA）、1,10-菲咯啉和金属蛋白酶组织抑制剂（TIMP）抑制；（2）这种活性与明胶-琼脂糖亲和树脂结合；（3）在十二烷基硫酸钠-聚丙烯酰胺凝胶（SDS-聚丙烯酰胺凝胶）上其质量约为92 kDa；（4）它能切割明胶；（5）这种可诱导的蛋白酶与抗MMP-9抗血清发生交叉反应。
