摘要目的 体外探讨凝血因子Ⅶa促进结肠癌细胞株SW620增殖与迁移的作用机制.方法 采用蛋白酶激活受体2激动剂(PAR2-AP)、凝血因子Ⅶa等处理SW620细胞,以实时定量PCR检测SW620细胞中白细胞介素8(IL-8)、组织因子(TF)及半胱氨酸蛋白酶7(caspase-7)mRNA的表达水平;以酶联免疫吸附试验(ELISA)检测细胞上清IL-8蛋白的含量;以Xa生成法检测细胞TF活性;以Western blot法检测细胞磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)水平.结果 PAR2-AP、凝血因子Ⅶa能够增加SW620细胞中IL-8基因和蛋白的表达,上调TF mRNA水平及活性,下调caspase-7基因表达和p-p38 MAPK水平,单克隆抗TF及抗PAR2抗体均可抑制凝血因子Ⅶa的作用.结论 凝血因子Ⅶa与细胞表面TF形成复合物,通过活化PAR2,上调结肠癌细胞株SW620中IL-8、TF的表达,下调细胞caspase-7表达,从而促进细胞增殖与迁移能力,p38 IVIAPK在此过程中起负性调节作用.

abstractsObjective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process.