Sakaki T, Fujita A, Wakumoto N, Murase S
Gan. 1976 Feb;67(1):67-73.
In order to demonstrate a collagenolytic enzyme in a tumor, solid Tawa sarcoma was subjected to a tissue culture technique. Salt- and acid-soluble collagen used as substrate was extracted from the rat skin and tail tendon. The tumor mass used was obtained on the 5th, 8th, and 11th days after subcutaneous transplantation of ascites Tawa sarcoma cells. Each tissue fragment in the inner and outer layers of the tumor mass was incubated on collagen gel at 37 degrees for 4 days, and collagenolytic activity was determined by comparing the relative content of hydroxyproline in the attacked collagen which was separated by means of centrifugation. All fragments were found to possess collagenolytic activity with some variation. Higher activity was observed in the outer layer than in the inner layer. Tumor fragments were cultured, and the collagenolytic enzyme was isolated from the culture medium and concentrated by ammonium sulfate precipitation and acrylamide disc electrophoresis. Collagenolytic enzyme activity was examined for its mode of attack on native collagen.
为了在肿瘤中证明一种胶原分解酶，对实体塔瓦肉瘤采用了组织培养技术。用作底物的盐溶性和酸溶性胶原是从大鼠皮肤和尾腱中提取的。所用肿瘤块是在腹水型塔瓦肉瘤细胞皮下移植后的第5天、第8天和第11天获得的。将肿瘤块内层和外层的每个组织碎片在37摄氏度下于胶原凝胶上孵育4天，并通过比较经离心分离的被侵袭胶原中羟脯氨酸的相对含量来测定胶原分解活性。发现所有碎片都具有胶原分解活性，但存在一些差异。观察到外层的活性高于内层。培养肿瘤碎片，从培养基中分离出胶原分解酶，并通过硫酸铵沉淀和丙烯酰胺圆盘电泳进行浓缩。检测胶原分解酶对天然胶原的攻击模式。
