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目的探讨凝血因子Ⅶa对结肠癌SW620细胞增殖能力以及表达凋亡分子天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)的影响及其作用机制。方法用一定剂量的蛋白酶激活受体2激动剂(PAR2-AP)、凝血因子Ⅶa等刺激物处理SW620细胞,观察细胞生长情况;western blot和定量PCR分别检测细胞表达caspase-3蛋白质及mRNA水平;利用相关抗体、拮抗剂和抑制剂等观察因子Ⅶa对caspase-3表达的效应变化。结果 PAR2-AP(100μmol/L)及因子Ⅶa(10 nmol/L)能够明显促进SW620细胞的生长,减少细胞caspase-3蛋白质和mRNA的表达;抗组织因子(TF)抗体(а-TF)、PAR2拮抗剂(PAR2-аAP)、ERK1/2抑制剂U0126以及NF-κB抑制剂PDTC均能逆转因子Ⅶa对SW620细胞caspase-3表达的抑制效应,而p38MAPK抑制剂SB203580对caspase-3的表达无明显的干预作用。结论因子Ⅶa依赖TF活化PAR2,经ERK1/2和NF-κB信号通路,抑制SW620细胞caspase-3表达,从而促进细胞的增殖与生长。
Objective To investigate the effect of coagulation factor Ⅶ a on the proliferation of colon cancer SW620 cells and the expression of caspase-3 and its mechanism of action. Methods SW620 cells were treated with a certain dose of PAR2-AP and coagulation factor Ⅶ a to observe the cell growth. Western blot and quantitative PCR were used to detect the expression of caspase-3 protein and mRNA. The effect of factor Ⅶ a on the expression of caspase-3 was observed by using relevant antibodies, antagonists and inhibitors. Results PAR2-AP (100μmol / L) and factor Ⅶa (10 nmol / L) could significantly promote the growth of SW620 cells and decrease the expression of caspase-3 protein and mRNA. Anti- PAR2 antagonist (PAR2-аAP), ERK1 / 2 inhibitor U0126 and NF-κB inhibitor PDTC could reverse the inhibitory effect of factor Ⅶa on the expression of caspase-3 in SW620 cells, while p38MAPK inhibitor SB203580 had no effect on the expression of caspase-3 Obvious intervention. Conclusion Factor Ⅶ a dependent on TF activates PAR2 and inhibits the expression of caspase-3 in SW620 cells via ERK1 / 2 and NF-κB signaling pathways, thereby promoting cell proliferation and growth.