Clauss M, Murray J C, Vianna M, de Waal R, Thurston G, Nawroth P, Gerlach H, Bach R, Familletti P C, Stern D
Department of Physiology and Cellular Biophysics, Columbia University, New York, New York 10032.
J Biol Chem. 1990 Apr 25;265(12):7078-83.
Intravascular clot formation, localized to the neoplasm, is an early component of the vascular response to tumor necrosis factor (TNF)/cachectin. Fibrin is closely associated with the endothelial cell surface, and multiple microthromboses lead to reduced blood flow in the tumor. We have identified a tumor-derived mediator which enhances endothelial procoagulant activity and the cellular response to TNF using cultured cells derived from a murine methylcholanthrene A (meth A)-induced fibrosarcoma as a model system. A heat-stable protease K-sensitive polypeptide, Mr approximately 44,000 on nonreduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mr approximately 56,000 reduced), was purified approximately 500,000-fold from serum-free culture supernatants of meth A cells by sequential Q-Sepharose, Mono S, reversed phase, and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Based on immunologic criteria, biologic activity, and other molecular properties, meth A factor appears to be distinct from other cytokines and growth factors. Purified meth A factor induced transcription of the tissue factor gene and expression of procoagulant activity by cultured human endothelium (half-maximal effect for the latter at approximately 6-8 pM). Furthermore, co-incubation of endothelium with meth A factor together with TNF enhanced induction of tissue factor in a more than additive manner. These data indicate that certain tumors elaborate an apparently unique molecule which can alter hemostatic properties of the vessel wall, potentially modulating reactivity of the tumor vasculature to host response mediators.
局限于肿瘤内的血管内血栓形成是血管对肿瘤坏死因子（TNF）/恶病质素反应的早期组成部分。纤维蛋白与内皮细胞表面密切相关，多个微血栓形成导致肿瘤内血流减少。我们使用源自小鼠甲基胆蒽A（meth A）诱导的纤维肉瘤的培养细胞作为模型系统，鉴定出一种肿瘤衍生介质，它可增强内皮促凝活性以及细胞对TNF的反应。一种热稳定的蛋白酶K敏感多肽，在非还原十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上的分子量约为44,000（还原后约为56,000），通过连续的Q-琼脂糖凝胶、单克隆S、反相和制备型十二烷基硫酸钠-聚丙烯酰胺凝胶电泳从meth A细胞的无血清培养上清液中纯化了约500,000倍。基于免疫学标准、生物学活性和其他分子特性，meth A因子似乎与其他细胞因子和生长因子不同。纯化的meth A因子可诱导培养的人内皮细胞组织因子基因的转录和促凝活性的表达（后者的半最大效应约为6 - 8 pM）。此外，内皮细胞与meth A因子以及TNF共同孵育可增强组织因子的诱导，且呈超加性方式。这些数据表明某些肿瘤会分泌一种明显独特的分子，它可改变血管壁的止血特性，潜在地调节肿瘤血管对宿主反应介质的反应性。
