Ogiichi T, Hirashima Y, Nakamura S, Endo S, Kurimoto M, Takaku A
Department of Neurosurgery, Toyama Medical and Pharmaceutical University, Japan.
J Neurooncol. 2000;46(1):1-9. doi: 10.1023/a:1006323200001.
The relationship between coagulation cascade activation and glioma cell proliferation was examined. The human glioma cell lines T98G, TM-1 and normal human astrocyte cell strain (NHA) were examined. Using anti-tissue factor (TF) antibody, immunocytochemical detection of TF antigen was obtained in both cell lines and cell strain. TF antigen in cell lysates was also measured by enzyme linked immunosorbent assay (ELISA). In a one-stage clotting assay, T98G, TM-1 and NHA revealed procoagulant activity (PCA) in normal human plasma and factor VII deficient plasma. PCA in normal human plasma was significantly inhibited by both inhibitory anti-TF antibody and cysteine protease inhibitor HgCl2. This result indicates that T98G, TM-1 and NHA cells express not only TF but also cancer procoagulant (CP) at the same time. In a cell proliferation assay, thrombin induced proliferation in T98G and TM-1 cells in a dose-dependent fashion and in NHA cell in a bell-shaped fashion. This mitogenic stimulant was inhibited by the specific thrombin inhibitor hirudin. The combinations of coagulation factors II, V, and X with or without factor VII induced proliferation in T98G, TM-1, and NHA cells. The maximal mitogenic stimulatory effects were larger in glioma cells than in NHA. These mitogenic stimulatory effects were also inhibited by hirudin. Each coagulation factor on its own or in any other combination of coagulation factors had no proliferative effect. Thus, these mitogenic stimulatory effects were considered to be the effect of thrombin. In conclusion, T98G and TM-1 human glioma cells express two different types of procoagulants TF and CP. In the presence of coagulation factors, these glioma cells can generate thrombin and this thrombin generation is capable of inducing glioma cell proliferation in vitro.
研究了凝血级联激活与胶质瘤细胞增殖之间的关系。检测了人胶质瘤细胞系T98G、TM-1和正常人星形胶质细胞株（NHA）。使用抗组织因子（TF）抗体，通过免疫细胞化学检测在两种细胞系和细胞株中均获得了TF抗原。细胞裂解物中的TF抗原也通过酶联免疫吸附测定（ELISA）进行了测量。在一期凝血试验中，T98G、TM-1和NHA在正常人血浆和因子VII缺乏血浆中均显示出促凝活性（PCA）。正常人血浆中的PCA被抑制性抗TF抗体和半胱氨酸蛋白酶抑制剂HgCl2均显著抑制。该结果表明T98G、TM-1和NHA细胞同时不仅表达TF，还表达癌促凝剂（CP）。在细胞增殖试验中，凝血酶以剂量依赖性方式诱导T98G和TM-1细胞增殖，并以钟形方式诱导NHA细胞增殖。这种促有丝分裂刺激物被特异性凝血酶抑制剂水蛭素抑制。凝血因子II、V和X与或不与因子VII的组合诱导T98G、TM-1和NHA细胞增殖。胶质瘤细胞中的最大促有丝分裂刺激作用大于NHA细胞。这些促有丝分裂刺激作用也被水蛭素抑制。每种凝血因子单独或与任何其他凝血因子组合均无增殖作用。因此，这些促有丝分裂刺激作用被认为是凝血酶的作用。总之，T98G和TM-1人胶质瘤细胞表达两种不同类型的促凝剂TF和CP。在凝血因子存在的情况下，这些胶质瘤细胞可产生凝血酶，并且这种凝血酶的产生能够在体外诱导胶质瘤细胞增殖。
