凝血酶促进结肠癌SW480细胞生长的研究

目的探讨凝血酶对结肠癌SW480细胞增殖及细胞周期的影响。方法将20 nmol/L、100 nmol/L、500nmol/L的凝血酶作用于SW480细胞48 h,以未处理组（0 nmol/L）作为对照。CCK-8（cell counting kit-8）法检测细胞增殖情况;流式细胞术检测细胞周期变化;RT-PCR检测蛋白酶活化受体-4（protease-activated receptor-4,PAR-4）的mRNA表达;Western blotting检测NF-κB p50蛋白表达。结果在20-500 nmol/L浓度范围内,凝血酶可剂量依赖性的促进SW480细胞的增殖;细胞周期显示G0/G1期细胞百分数减少,S期及G2/M期细胞百分数增加,表明凝血酶可诱导细胞从G0/G1期向G2/M期发展,从而促进细胞增殖;随着凝血酶浓度的增加,PAR-4 mRNA及NF-κB p50蛋白含量也有不同程度的增加。结论在20-500 nmol/L浓度范围内,凝血酶可促进结肠癌SW480细胞的增殖,加速细胞周期进程;其作用可经PAR-4介导,并与NF-κB信号通路活化有关。

Study of the role of thrombin on facilitation the growth of colon carcinoma SW480 cells

Objective To investigate the effects of thrombin on cell proliferation and cell cycle of colon carcinoma SW480 cells.Methods SW480 cells were treated with thrombin by three different concentration,20 nmol/L,100 nmol/L,500 nmol/L,and untreated group（0 nmol/L） as a control.Cell proliferation activity was analysed by cell counting kit-8,cell cycle was detected by flow cytometry,the mRNA of protease-activated receptor-4（PAR-4） was measured by RT-PCR,and the protein of NF-κB p50 was examined by Western blotting.Results At the scope of 20-500 nmol/L,thrombin could facilitate the growth of SW480 cells with the dose-dependent manner.Cell cycle analysis showed that the percent of G0/G1 phase cell was significantly decreased,S and G2/M phase cell were significantly increased,which suggested that thrombin could induce cell proliferation by increasing the number of cells from G0/G1 phase to G2/M phase.The expression of PAR-4 mRNA and NF-κB p50 was also increased in different levels along with the increasing concentration of the thrombin.Conclusion Thrombin can facilitate the growth of colon carcinoma SW480 cells,and accelerate the process of cell cycle,which occurrence may be mediated by PAR-4,and may involve in the activation of NF-κB pathway.