In vitro inhibition of human sarcoma cells' invasive ability by bis(5-amidino-2-benzimidazolyl)methane--a novel esteroprotease inhibitor.
In vitro inhibition of human sarcoma cells' invasive ability by bis(5-amidino-2-benzimidazolyl)methane--a novel esteroprotease inhibitor.
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双(5-脒基-2-苯并咪唑基)甲烷——一种新型酯蛋白酶抑制剂对人肉瘤细胞侵袭能力的体外抑制。
DOI：
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发表时间：
1986
影响因子：
6
通讯作者：
G. Siegal
中科院分区：
文献类型：
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作者：
D. H. Cresson;W. Beckman;R. Tidwell;J. Geratz;G. Siegal
双（5-脒基-2-苯并咪唑基）甲烷（BABIM）是一种合成的芳香族脒化合物，其具有许多重要的生物化学作用，包括抑制先前与肿瘤侵袭的复杂过程相关的酯蛋白酶家族（胰蛋白酶、尿激酶、纤溶酶）。以往的工作表明，外源性天然蛋白酶抑制剂可以在体外阻断肿瘤细胞穿过基底膜（BM）的侵袭。作者研究了BABIM对人细胞系HT-1080的作用，使用体外羊膜侵袭定量分析系统。他们已经验证了这些细胞在裸鼠中生长的能力，并根据接种物的给药途径通过血管或血管转移。将能够主动穿过整个BM的细胞捕获在过滤器上，并通过明视野显微镜和其DNA用氚化胸苷标记的细胞的β闪烁计数进行计数。与两种计数技术一致，在7天的实验过程中，浓度为10（-4）M的BABIM显著抑制侵袭（P <0.005）。在这些条件下，抑制剂是无毒的，并且不改变细胞与羊膜的附着。此外，在超过2个数量级的BABIM的摩尔浓度变化中也证明了对侵袭的高度显著的抑制（P小于0.001）。最显著的是，在10（-9）和10（-3）M BABIM存在下，细胞的侵袭能力最初受到抑制。对来自这些细胞的培养基中的IV型特异性胶原酶的测量显示在BABIM存在下活性的显著抑制。这些结果表明两个，不一定排他的，替代的解释：第一，抑制蛋白水解步骤沿着活化的基底膜降解酶的途径导致抑制入侵;第二，精氨酸指导的酯蛋白酶可能与细胞的胶原溶解金属蛋白酶在人类肿瘤细胞通过天然基质屏障的入侵过程中协同工作。
Bis(5-amidino-2-benzimidazolyl)methane (BABIM) is a synthetic aromatic amidine compound which has a number of important biochemical effects, including inhibition of a family of esteroproteases (trypsin, urokinase, plasmin) previously linked to the complex process of tumor invasion. Previous work has suggested that exogenous natural protease inhibitors can block invasion of tumor cells across basement membranes (BM) in vitro. The authors studied the effect of BABIM on the human cell line HT-1080 with the use of a quantitative in vitro amnion invasion assay system. They have verified the ability of these cells to grow in nude mice and metastasize via the lymphatics or blood vessels on the basis of the route of administration of the inoculum. Cells which were able to actively cross the entire BM were trapped on filters and counted by both brightfield microscopy and by beta scintillation counting of cells whose DNA was labeled with tritiated thymidine. In agreement with either counting technique, BABIM, at a concentration of 10(-4) M, significantly inhibited invasion (P less than 0.005) over the 7-day course of the experiments. Under these conditions, the inhibitor was nontoxic and did not alter the attachment of the cells to the amniotic membrane. Furthermore, a highly significant inhibition of invasion (P less than 0.001) was also demonstrated across a variation in molar concentration of BABIM of more than 2 orders of magnitude. Most remarkably, cells were initially inhibited in their ability to invade in the presence of between 10(-9) and 10(-3) M BABIM. Measurement of Type IV specific collagenase in media from these cells shows a significant inhibition of activity in the presence of BABIM. These results suggest two, not necessarily exclusive, alternative interpretations: first, that inhibition of the proteolytic steps along the pathway of activation of basement membrane degrading enzymes results in inhibition of invasion; second, that arginine directed esteroproteases may work in concert with cellular collagenolytic metalloproteinases in the process of invasion by human tumor cells through native matrix barriers.