Nierodzik M L, Kajumo F, Karpatkin S
New York University Medical School, New York 10016.
Cancer Res. 1992 Jun 15;52(12):3267-72.
Seven different tumor cell lines (human melanoma SK MEL 28; hamster melanoma HM29; murine melanomas B16F10 and amelanotic melanoma B16a; human colon carcinoma HCT8; murine colon carcinoma CT26; and murine Lewis lung carcinoma) were treated with thrombin at 0.5-1 unit/ml and examined for their ability to bind to adherent platelets; HM29 was studied for its ability to bind to fibronectin and von Willebrand factor; CT26, B16F1, B16F10, and B16a were studied for their ability to form pulmonary metastasis after i.v. injection of thrombin-treated tumor cells; CT26 was studied for its ability to grow s.c. Five of 7 thrombin-treated tumor cell lines increased their adhesion to adherent platelets 2-to 3-fold. HM29 increased its adherence to fibronectin and von Willebrand factor 2- to 3-fold. CT26, B16F1, B16F10, and B16a increased experimental pulmonary metastasis 10- to 156-fold. Thrombin-treated CT26 cells demonstrated 2-fold greater growth in vivo after s.c. injection. The mechanism of enhanced adhesion of thrombin-treated tumor cells to platelets required the platelet integrin GPIIb-GPIIIa since it could be inhibited by agents known to block adhesion of ligands to GPIIb-GPIIIa (monoclonal antibody 10E5, tetrapeptide RGDS, disintegrin Albolabrin); as well as a "GPIIb-GPIIIa-like" structure on tumor cells since it could be inhibited by treatment of thrombin-treated tumor cells with 10E5 and RGDS. The thrombin effect on tumor cells was optimum at 1 h of incubation with thrombin, did not require active thrombin on the tumor cell surface, and did not require protein synthesis (not inhibited by cycloheximide). Thus, thrombin-treated tumor cells markedly enhance pulmonary metastasis. It is suggested that this may be secondary to thrombin-induced enhanced adhesion as well as growth of tumor cells.
七种不同的肿瘤细胞系（人黑色素瘤SK MEL 28；仓鼠黑色素瘤HM29；小鼠黑色素瘤B16F10和无黑色素黑色素瘤B16a；人结肠癌HCT8；小鼠结肠癌CT26；以及小鼠Lewis肺癌）用0.5 - 1单位/毫升的凝血酶处理，并检测它们与黏附血小板结合的能力；研究了HM29与纤连蛋白和血管性血友病因子结合的能力；研究了CT26、B16F1、B16F10和B16a经静脉注射凝血酶处理的肿瘤细胞后形成肺转移的能力；研究了CT26皮下生长的能力。7种经凝血酶处理的肿瘤细胞系中有5种与黏附血小板的黏附能力增加了2至3倍。HM29与纤连蛋白和血管性血友病因子的黏附增加了2至3倍。CT26、B16F1、B16F10和B16a的实验性肺转移增加了10至156倍。经凝血酶处理的CT26细胞皮下注射后在体内的生长速度提高了2倍。凝血酶处理的肿瘤细胞与血小板黏附增强的机制需要血小板整合素GPIIb - GPIIIa，因为它可被已知能阻断配体与GPIIb - GPIIIa黏附的试剂（单克隆抗体10E5、四肽RGDS、去整合素Albolabrin）抑制；肿瘤细胞上还需要一种“GPIIb - GPIIIa样”结构，因为用10E5和RGDS处理凝血酶处理的肿瘤细胞可抑制该机制。凝血酶对肿瘤细胞的作用在与凝血酶孵育1小时时最为显著，不需要肿瘤细胞表面有活性凝血酶，也不需要蛋白质合成（不受环己酰亚胺抑制）。因此，凝血酶处理的肿瘤细胞显著增强了肺转移。提示这可能继发于凝血酶诱导的肿瘤细胞黏附增强以及生长。
