Expression of a Mr 41,000 glycoprotein associated with thrombin-independent platelet aggregation in high metastatic variants of murine B16 melanoma.
Expression of a Mr 41,000 glycoprotein associated with thrombin-independent platelet aggregation in high metastatic variants of murine B16 melanoma.
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在鼠 B16 黑色素瘤的高转移变异体中，Mr 41,000 糖蛋白的表达与凝血酶独立的血小板聚集相关。
作者：
M. Watanabe;Y. Sugimoto;T. Tsuruo
在之前的研究中，我们制备了针对NL-17的单抗8F11，NL-17是来自小鼠结肠腺癌26的转移变体的高转移克隆。8F11可抑制NL-17诱导的血小板聚集，并识别MR 44000膜蛋白为抗原。本研究检测了8F11对小鼠B16黑色素瘤及其转移株的反应性，并对8F11识别的细胞表面抗原进行了鉴定。8F11与B16黑色素瘤的3种转移变异体呈强阳性反应。相比之下，在亲本B16黑色素瘤中只观察到轻微的反应。抗体对这些细胞的反应性为B16F10>B16BL-6>B16F1，远大于B16。Western印迹分析表明，B16F10细胞表面的MR 41000蛋白是8F11识别的抗原。MR 41000抗原似乎是一种与小麦胚芽凝集素结合的糖蛋白，正如对NL-17的MR 44000抗原所观察到的那样。为了阐明MR 41000抗原在B16黑色素瘤中的功能作用，比较了B16和B16F10诱导的血小板聚集。据报道，B16通过凝血酶的生成刺激血小板聚集，而B16F10至少通过两种机制激活血小板：一种依赖于凝血酶，另一种不依赖凝血酶。在人工合成的凝血酶拮抗剂MD805存在下，B16及其转移变异体诱导血小板聚集的活性与8F11对这些细胞的反应性很好地相关。8F11在防止凝血酶活性的条件下，如通过磷脂酶A2处理细胞表面或在MD805存在的情况下酶促形成溶血磷脂，可阻断B16F10对血小板的激活。这些数据表明，8F11识别的B16黑色素瘤转移变异体上的MR 41,000糖蛋白参与了凝血酶非依赖性的血小板聚集。B16及其转移变异体的MR 41000糖蛋白表达水平与静脉注射后肺转移呈正相关。注射，提示MR 41,000糖蛋白，以及之前报道的其他因素，可能在B16黑色素瘤的血源性扩散中发挥重要作用。
In the previous study, we generated a monoclonal antibody, 8F11, against NL-17, a high metastatic clone derived from a metastatic variant of murine colon adenocarcinoma 26. 8F11 inhibited platelet aggregation induced by NL-17 and recognized a Mr 44,000 membrane protein as antigen. In the present study, the reactivity of 8F11 to murine B16 melanoma and its metastatic variants was examined, and the antigen recognized by 8F11 on the cell surface was characterized. 8F11 was found to strongly react with 3 metastatic variants of B16 melanoma. In contrast, only slight reactivity was observed with parent B16 melanoma. The reactivity of the antibody to these cells was in the order B16F10 greater than B16BL-6 greater than B16F1 much greater than B16. Western blot analysis showed a Mr 41,000 protein as the antigen recognized by 8F11 on the cell surface of B16F10 cells. The Mr 41,000 antigen appeared to be a glycoprotein that bound to wheat germ agglutinin as has been observed for the Mr 44,000 antigen of NL-17. To elucidate the functional role of the Mr 41,000 antigen in B16 melanoma, platelet aggregation induced by B16 and B16F10 was compared. B16 was reported to stimulate platelet aggregation by the generation of thrombin, whereas B16F10 was found to activate platelet by at least 2 mechanisms: one dependent on thrombin and the other independent on thrombin. The activity of B16 and its metastatic variants to induce platelet aggregation in the presence of MD805, a synthetic antagonist of thrombin, well correlated with the reactivity of 8F11 to these cells. 8F11 blocked platelet activation by B16F10 under conditions preventing thrombin activity such as enzymatic formation of lysolecithin through the treatment of the cell surface with phospholipase A2 or in the presence of MD805. These data indicate that Mr 41,000 glycoprotein recognized by 8F11 on metastatic variants of B16 melanoma is involved in the thrombin-independent platelet aggregation. A positive correlation was observed between the levels of Mr 41,000 glycoprotein expression of B16 and its metastatic variants and their pulmonary metastasis after i.v. injection, suggesting Mr 41,000 glycoprotein, as well as other factors reported previously, may play an important role in the hematogenous spread of B16 melanoma.
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DOI：
10.1073/pnas.85.2.612
发表时间：
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影响因子：
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作者：
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影响因子：
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