【摘要】 目的:探讨细胞外信号调节激酶1/2(extracellular signal-regulated kinase1/2,ERK1/2)/钙激活中性蛋白酶(calcium-activated neutral protease,CANP)快速(非基因组)信号通路参与介导17β-雌二醇(17β-estradiol,E2)诱导人乳腺癌MCF-7细胞的增殖。方法:用E2处理MCF-7细胞,需要时以ERK1/2阻断剂PD98059或CANP特异性抑制剂calpeptin预处理MCF-7细胞,应用MTT法检测MCF-7细胞的增殖改变,FCM法检测细胞周期变化,Western印迹法检测细胞总ERK、p-ERK和CANP小亚基Capn4蛋白的表达水平。结果:10-8mol/LE2作用24h可使MCF-7细胞显著增殖(P<0.01),G0/G1期细胞减少,G1/S期转换加速;Calpeptin可显著抑制E2诱导的MCF-7细胞增殖效应(P<0.01),G0/G1期细胞增多,细胞延缓进入S期。E2可快速(10min)诱导MCF-7细胞p-ERK表达上调并持续16h(P<0.01),但对总ERK表达水平无明显影响,PD98059对E2诱导的MCF-7细胞ERK磷酸化具有显著抑制作用(P<0.01)。E2可快速(10min)刺激MCF-7细胞Capn4蛋白表达上调(P<0.01),PD98059或Calpeptin对E2诱导的Capn4蛋白表达均有显著抑制作用(P<0.01)。结论:E2可通过ERK/CANP快速信号通路诱导乳腺癌细胞增殖。更多还原

【Abstract】 Objective:To investigate a possible role of extracellular signal-regulated kinase(ERK) 1/2/calcium-activated neutral protease(CANP) signaling pathway in 17β-estradiol(E2)-induced proliferation of human breast cancer cell line MCF-7.Methods:MCF-7 cells were pretreated with ERK1/2 inhibitor PD98059 or CANP inhibitor calpeptin,and then treated with 1×10-8 mol/L E2.MTT assay was employed to determine the effect on cell proliferation.Cell cycle distribution was detected by flow cytometry(FCM).The expression levels of phosphorylated-ERK(p-ERK),total ERK and CANP small subunit Capn4 proteins were examined by Western blotting.Results:The proliferation rate was increased in MCF-7 cells treated with 1×10-8 mol/L E2 for 24 h(P<0.01).The number of cells in G0/G1 phase was decreased,and the accelerated G1/S transition was observed.Calpeptin could significantly inhibited E2-induced proliferation of MCF-7 cells(P<0.01).The number of cells in G0/G1 phase was increased,and the delayed G1/S transition was observed.The expression of p-ERK was rapidly up-regulated(during 10 minutes) induced by E2(P<0.01),and this effect persisted for at least 16 h;but there was no obvious change in the expression of total ERK.E2-induced ERK phosphorylation in MCF-7 cells was significantly inhibited by PD98059(P<0.01).The expression of Capn4 protein was rapidly up-regulated in E2-induced MCF-7 cells(P<0.01) while significantly inhibited by PD98059 or calpeptin(P<0.01) in MCF-7 cells.Conclusion:The rapid signaling pathway of ERK/CANP is involved in E2-induced proliferation of human breast cancer cells.更多还原